Pharmaceutical compositions comprising dgla and use of same

ABSTRACT

The present disclosure provides orally deliverable pharmaceutical compositions comprising DGLA and to methods of using same to treat a variety of conditions and disorders.

TECHNICAL FIELD

The present application relates generally to pharmaceutical compositionscomprising DGLA and methods of using same.

BACKGROUND

Dihomo gamma linolenic acid (DGLA) is an essential fatty acid foundnaturally in the body as the elongation product of gamma linolenic acid(GLA). GLA is in turn a desaturation product of linoleic acid. Softgelatin encapsulation of DGLA is challenging as it is prone to oxidationto aldehydes which can interact with amino groups in the gelatin polymerin the capsule shell. This can cause slowdown in drug release due tocrosslinking of the gelatin polymers.

BRIEF DESCRIPTION OF THE DRAWINGS

Many aspects of the present disclosure can be better understood withreference to the following drawings. The components in the drawings arenot necessarily to scale. Instead, emphasis is placed on illustratingclearly the principles of the present disclosure.

FIG. 1 shows change in mean arterial pressure (mm Hg) with intravenousdoses of phenylephrine following seven consecutive days of gavage withAspirin at 10 mg/kg/day.

FIG. 2 shows change in mean arterial pressure (mmHg) with intravenousdoses of phenylephrine following seven consecutive days of gavage withDGLA at 50 mg/kg+Aspirin at 10 mg/kg.

FIG. 3 shows change in mean arterial pressure (mm Hg) with intravenousdoses of phenylephrine following seven consecutive days of gavage withDGLA at 500 mg/kg co-administered with Aspirin at 10 mg/kg.

FIG. 4 shows mean arterial pressure at baseline following sevenconsecutive days with six different gavage groups.

FIG. 5 shows mean arterial pressure with an intravenous dose ofphenylephrine at 20 μk/kg following seven consecutive days with sixdifferent gavage groups.

FIG. 6 shows mean plasma free DGLA concentration (ng/mL, linear plot),by dose cohort (Single Dose, PK Population).

FIG. 7 shows mean plasma free DGLA concentration (ng/mL, log-linearplot), by dose cohort (Single Dose, PK Population).

FIG. 8 shows mean plasma total DGLA concentration (ng/mL, linear plot),by dose cohort (Single Dose, PK Population).

FIG. 9 shows mean plasma total DGLA concentration (ng/mL, log-linearplot), by dose cohort (Single Dose, PK Population).

FIG. 10 shows mean plasma free DGLA concentration (ng/mL, linear plot),by dose cohort (Multiple-dose, PK Population).

FIG. 11 shows mean plasma free DGLA Concentration (ng/mL, log-linearplot), by dose cohort (Multiple-dose, PK Population).

FIG. 12 shows mean plasma total DGLA concentration (ng/mL, linear plot),by dose cohort (Multiple-dose, PK Population).

FIG. 13 shows mean plasma total DGLA concentration (ng/mL, log-linearplot), by dose cohort (Multiple-dose, PK Population).

FIG. 14 shows mean skin blister fluid concentration of Free DGLA (ng/mL,linear plot), by dose cohort (Multiple-dose, PK Population).

FIG. 15 shows mean skin blister fluid concentration of free DGLA (ng/mL,log-linear plot), by dose cohort (Multiple-dose, PK Population).

FIG. 16 shows mean skin blister fluid concentration of total DGLA(ng/mL, linear plot), by dose cohort (Multiple-dose, PK Population).

FIG. 17 shows mean skin blister fluid concentration of total DGLA(ng/mL, log-linear plot), by dose cohort (Multiple-dose, PK Population)

FIG. 18 shows mean free DGLA concentration (ng/mL, linear plot) inplasma and skin blister fluid, by dose cohort (Multiple-dose, PKPopulation).

FIG. 19 shows mean free DGLA concentration (ng/mL, log-linear plot) inplasma and skin blister fluid, by dose cohort (Multiple-dose, PKPopulation).

FIG. 20 shows mean total DGLA concentration (ng/mL, linear plot) inplasma and skin blister fluid, by dose cohort (Multiple-dose, PKPopulation).

FIG. 21 shows mean total DGLA concentration (ng/mL, log-linear plot) inplasma and skin blister fluid, by dose cohort (Multiple-dose, PKPopulation).

FIG. 22 shows mean plasma dihydrotestosterone concentration (ng/mL,linear plot), by dose cohort (Multiple-dose, PK Population).

FIG. 23 shows mean plasma dihydrotestosterone concentration (ng/mL,log-linear plot), by dose cohort (Multiple-dose, PK Population).

SUMMARY

The present disclosure provides orally deliverable pharmaceuticalcompositions comprising DGLA and to methods of using same to treat avariety of conditions and disorders.

In one embodiment, the present disclosure provides a pharmaceuticalcomposition comprising DGLA. In one embodiment, the composition isencapsulated in a capsule shell. In one embodiment, the presentdisclosure provides a pharmaceutical composition comprising DGLAencapsulated in a capsule shell comprising gelatin, d-sorbitol and1,4-sorbitan sugar alcohols. In one embodiment, about 500 mg to about 1g of DGLA or a derivative thereof is encapsulated in the capsule shell.

In one embodiment, the present disclosure provides a method of treatinga skin disease or disorder in a subject in need thereof, the methodcomprising orally administering to the subject a pharmaceuticalcomposition comprising DGLA. Optionally, the pharmaceutical compositioncomprises DGLA encapsulated in a capsule shell comprising gelatin,d-sorbitol and 1,4-sorbitan sugar alcohols. Optionally, the compositionis administered to the subject in an amount sufficient to provide about1 g to about 4 g of DGLA per day. In one embodiment, the gelatin has agel mass viscosity of about 9500 to about 11000, for example about 9775or about 10,500. In another embodiment, the gelatin has a bloom of about165 to about 190, for example about 170 to about 185. In anotherembodiment, the gelatin has an ash percentage>than about 0.33.

In one embodiment, the present disclosure provides a method of treatingoveractive bladder in a subject in need thereof, the method comprisingorally administering to the subject a pharmaceutical compositioncomprising DGLA. Optionally, the pharmaceutical composition comprisesDGLA encapsulated in a capsule shell comprising gelatin, d-sorbitol and1,4-sorbitan sugar alcohols. Optionally, the composition is administeredto the subject in an amount sufficient to provide about 1 g to about 4 gof DGLA per day. In one embodiment, the gelatin has a gel mass viscosityof about 9500 to about 11000, for example about 9775 or about 10,500. Inanother embodiment, the gelatin has a bloom of about 165 to about 190,for example about 170 to about 185. In another embodiment, the gelatinhas an ash percentage>than about 0.33. These and other embodiments ofthe invention are described in further detail below.

DETAILED DESCRIPTION

While the present invention is capable of being embodied in variousforms, the description below of several embodiments is made with theunderstanding that the present disclosure is to be considered as anexemplification of the invention, and is not intended to limit theinvention to the specific embodiments illustrated. Headings are providedfor convenience only and are not to be construed to limit the inventionin any manner. Embodiments illustrated under any heading may be combinedwith embodiments illustrated under any other heading.

The use of numerical values in the various quantitative values specifiedin this application, unless expressly indicated otherwise, are stated asapproximations as though the minimum and maximum values within thestated ranges were both preceded by the word “about.” In this manner,slight variations from a stated value can be used to achievesubstantially the same results as the stated value. Also, the disclosureof ranges is intended as a continuous range including every valuebetween the minimum and maximum values recited as well as any rangesthat can be formed by such values. Also disclosed herein are any and allratios (and ranges of any such ratios) that can be formed by dividing arecited numeric value into any other recited numeric value. Accordingly,the skilled person will appreciate that many such ratios, ranges, andranges of ratios can be unambiguously derived from the numerical valuespresented herein and in all instances such ratios, ranges, and ranges ofratios represent various embodiments of the present invention.

Compositions

In various embodiments, the present disclosure provides orallydeliverable pharmaceutical compositions comprising DGLA or a derivativethereof. The term DGLA herein refers to DGLA in free acid form.Compositions of the invention may also comprise a DGLA derivative inaddition to or instead of DGLA. Such derivatives include alkyl esters,lower alky esters such as DGLA methyl or ethyl ester or DGLA intriglyceride form. In one embodiment, the present disclosure provides apharmaceutical composition comprising DGLA or derivative thereofencapsulated in a capsule shell. In one embodiment, about 500 mg toabout 1 g of DGLA or derivative thereof is encapsulated in the capsuleshell.

In one embodiment, the capsule shell comprises gelatin, for exampleGelatin RXL or lime bone gelatin with a lower molecular weight. Inanother embodiment, the capsule shell comprises Gelatin RXL that hasbeen treated by proteolytic enzyme to cut the gelatin pattern andeffectively decrease its molecular weight. In another embodiment, thepharmaceutical composition comprises DGLA esters of D-Sorbitol and1,4-sorbitan. In one embodiment, the capsule shell comprises (a) gelatinand (b) plasticizers selected from one or more of d-sorbitol and1,4-sorbitans. In one embodiment, the gelatin is as described in U.S.Pat. No. 7,485,323, hereby incorporated by reference herein in itsentirety.

In one embodiment, the plasticizer comprises 1-4 sorbitans in an amountfrom 20%-30%, for example about 24% and 28% (on a dry basis) and aD-sorbitol content of about 30%-50%, for example about 35% to 45% (on adry basis).

In some embodiments, the capsule shell further comprises glycerol,purified water, titanium dioxide, medium chain triglycerides andlecithin.

In various embodiments, DGLA or a derivative is present in a compositionof the invention in an amount of about 50 mg to about 5000 mg, about 75mg to about 2500 mg, or about 100 mg to about 1000 mg, for example about75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450mg, about 475 mg, about 500 mg, about 525 mg, about 550 mg, about 575mg, about 600 mg, about 625 mg, about 650 mg, about 675 mg, about 700mg, about 725 mg, about 750 mg, about 775 mg, about 800 mg, about 825mg, about 850 mg, about 875 mg, about 900 mg, about 925 mg, about 950mg, about 975 mg, about 1000 mg, about 1025 mg, about 1050 mg, about1075 mg, about 1100 mg, about 1025 mg, about 1050 mg, about 1075 mg,about 1200 mg, about 1225 mg, about 1250 mg, about 1275 mg, about 1300mg, about 1325 mg, about 1350 mg, about 1375 mg, about 1400 mg, about1425 mg, about 1450 mg, about 1475 mg, about 1500 mg, about 1525 mg,about 1550 mg, about 1575 mg, about 1600 mg, about 1625 mg, about 1650mg, about 1675 mg, about 1700 mg, about 1725 mg, about 1750 mg, about1775 mg, about 1800 mg, about 1825 mg, about 1850 mg, about 1875 mg,about 1900 mg, about 1925 mg, about 1950 mg, about 1975 mg, about 2000mg, about 2025 mg, about 2050 mg, about 2075 mg, about 2100 mg, about2125 mg, about 2150 mg, about 2175 mg, about 2200 mg, about 2225 mg,about 2250 mg, about 2275 mg, about 2300 mg, about 2325 mg, about 2350mg, about 2375 mg, about 2400 mg, about 2425 mg, about 2450 mg, about2475 mg, or about 2500 mg. In any such embodiment, the composition canfurther comprise DGLA esters of D-Sorbitol and 1,4-sorbitan.

In one embodiment, a composition of the invention contains not more thanabout 10%, not more than about 9%, not more than about 8%, not more thanabout 7%, not more than about 6%, not more than about 5%, not more thanabout 4%, not more than about 3%, not more than about 2%, not more thanabout 1%, or not more than about 0.5%, by weight of total fatty acids,of fatty acids other than DGLA.

In another embodiment, DGLA or a derivative thereof represents at leastabout 30%, about 40%, about 50%, at least about 60%, at least about 70%,at least about 80%, at least about 90%, at least about 95%, at leastabout 97%, at least about 98%, at least about 99%, or 100%, by weight,of all fatty acids present in a composition of the invention.

In one embodiment, a composition of the invention, when placed in astandard disintegration test, for example as set for in USP 2040(Disintegration and Dissolution of Dietary Supplements) with water asthe Medium, has a DGLA release rate less than about 60 minutes, lessthan about 50 minutes, less than about 40 minutes, less than about 30minutes, or less than 20 minutes after storage for about 1 month, about2 months or about 3 months at 40° C./75% RH.

In one embodiment, after storage for about 1 month, about 2 months,about 3 months or about 6 months at 40° C./75% RH, a composition of theinvention comprises less than about 5% DGLA esters by weight of allfatty acids, less than about 4% DGLA esters by weight of all fattyacids, less than about 3% DGLA esters by weight of all fatty acids, lessthan about 2% DGLA esters by weight of all fatty acids, or less thanabout 1% DGLA esters by weight of all fatty acids.

Methods

Any composition of the invention, including compositions describedherein above or compositions that can for formulated from combiningvarious embodiments of the present disclosure, can be used in treatmentor prevention of: skin disorders and diseases, including acne vulgaris,acne rosacea, atopic dermatitis, psoriasis, pruritus/itch, radiationprotection, dry skin, smooth skin, healthy skin, anti-aging, andphotoprotection; urinary disorders and diseases including bladdercancer, cystocele, hematuria, interstitial cystitis, neurogenic bladder,Peyronie's disease, prostate disease, incontinence, urinary tractinfection and vasicoureteral reflux; renal disease and disordersincluding kidney failure, acute kidney injury, chronic kidney disease,and polycystic kidney disease; rheumatic disease including ankylosingspondylitis, fibromyalgia, gout, infectious arthritis, lupus,osteoarthritis, polymyalgia rheumatic, psoriatic arthritis, reactivearthritis, rheumatoid arthritis, sclerodoma; respiratory disordersincluding inflammatory lung disease, respiratory tract infections,pleural cavity disease, pulmonary vascular disease, pneumonia, pulmonaryembolism, and lung cancer; and cardiovascular disorders including acutecardiac ischemic events, acute myocardial infarction, angina,arrhythmia, atrial fibrulation, atherosclerosis, arterial fibrillation,cardiac insufficiency, cardiovascular disease, chronic heart failure,chronic stable angina, congestive heart failure, coronary arterydisease, coronary heart disease, deep vein thrombosis, diabetes,diabetes mellitus, diabetic neuropathy, diastolic dysfunction insubjects with diabetes mellitus, edema, essential hypertension, eventualpulmonary embolism, fatty liver disease, heart disease, heart failure,homozygous familial hypercholesterolemia (HoFH), homozygous familialsitosterolemia, hypercholesterolemia, hyperlipidemia, hypertension,hypertriglyceridemia, metabolic syndrome, mixed dyslipidemia, moderateto mild heart failure, myocardial infarction, obesity management,paroxysmal atrial/arterial fibrillation/fibrulation/flutter, paroxysmalsupraventricular tachycardias (PSVT), particularly severe or rapid onsetedema, platelet aggregation, primary hypercholesterolemia, primaryhyperlipidemia, pulmonary arterial hypertension, pulmonary hypertension,recurrent hemodynamically unstable ventricular tachycardia (VT),recurrent ventricular arrhythmias, recurrent ventricular fibrillation(VF), ruptured aneurysm, sitisterolemia, stroke, supraventriculartachycardia, symptomatic atrial fibrillation/flutter, tachycardia,type-II diabetes, vascular disease, venous thromboembolism, ventriculararrhythmias, and other cardiovascular events.

The term “treatment” in relation a given disease or disorder, includes,but is not limited to, inhibiting the disease or disorder, for example,arresting the development of the disease or disorder; relieving thedisease or disorder, for example, causing regression of the disease ordisorder; or relieving a condition caused by or resulting from thedisease or disorder, for example, relieving, preventing or treatingsymptoms of the disease or disorder. The term “prevention” in relationto a given disease or disorder means: preventing the onset of diseasedevelopment if none had occurred, preventing the disease or disorderfrom occurring in a subject that may be predisposed to the disorder ordisease but has not yet been diagnosed as having the disorder ordisease, and/or preventing further disease/disorder development ifalready present.

In various embodiments, compositions of the invention are administeredin an amount sufficient to provide a daily DGLA dose of about 50 mg toabout 10000 mg, about 100 mg to about 7500 mg, or about 100 mg to about5000 mg, for example about 200 mg, about 300 mg, about 400 mg, about 500mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000mg, about 1100 mg, about 1200 mg, about 1300 mg, about 1400 mg, about1500 mg, about 1600 mg, about 1700 mg, about 1800 mg, about 1900 mg,about 2000 mg, about 2100 mg, about 2200 mg, about 2300 mg, about 2400mg, about 2500 mg about 2600 mg, about 2700 mg, about 2800 mg, about2900 mg, about 3000 mg, about 3100 mg, about 3200 mg, about 3300 mg,about 3400 mg, about 3500 mg, 3600 mg, about 3700 mg, about 3800 mg,about 3900 mg, about 4000 mg, about 4100 mg, about 4200 mg, about 4300mg, about 4400 mg, about 4500 mg, 4600 mg, about 4700 mg, about 4800 mg,about 4900 mg, about 5000 mg, about 5100 mg, about 5200 mg, about 5300mg, about 5400 mg, about 5500 mg of DGLA per day.

In one embodiment, the invention provides a method of treating atopicdermatitis, for example mild to moderate atopic dermatitis. In oneembodiment, the method comprises administering to a subject in need ofsuch treatment DGLA in an amount of about 500 mg to about 3 g per day,about 1 g to about 2.5 g per day, about 1 g per day or about 2 g perday. In one embodiment, the DGLA is administered to the subject dailyfor a period of at least about 2 weeks, at least about 4 weeks or atleast about 8 weeks. In a related embodiment, upon treatment inaccordance with the present invention, for example over a period ofabout 1 to about 12 weeks, about 1 to about 8 weeks, or about 1 to about4 weeks, the subject or subject group exhibits one or more of thefollowing outcomes:

(a) a reduction in eczema area and severity index (EASI) score relativeto baseline or placebo control;

(b) a reduction in percentage of area of an anatomical site affected byatopic dermatitis relative to baseline or control;

(c) a reduction in investigator's global assessment score relative tobaseline or placebo control;

(d) a reduction in intensity of erythema, edema/population,oozing/crusts, excoriation, lichenification and/or dryness relative tobaseline or placebo control;

(e) a reduction in erythema, edema/population, oozing/crusts,excoriation, lichenification and/or dryness relative to baseline orplacebo control;

(f) a reduction in body surface area (BSA) affected by atopic dermatitisrelative to baseline or placebo control;

(g) a reduction in loss of sleep relative to baseline or placebocontrol;

(h) a reduction in occurrence of pruitis (itch) relative to baseline orplacebo control;

(i) a reduction in severity of pruritis as an average of the prior threedays and/or nights on a visual analog scale;

(j) a reduction in SCORAD score relative to baseline or placebo control;

(k) an improved patient-oriented Eczema Measure (POEM) compared tobaseline or placebo control;

(l) a reduction in number of days in the preceding week in which thesubject reported that the skin was itchy due to eczema;

(m) a reduction in number of days in the preceding week in which thesubject reported that their sleep was disturbed due to their eczema;

(n) a reduction in number of days in the preceding week in which thesubject experienced skin bleeding;

(o) a reduction in number of days in the preceding week in which thesubject experienced skin weeping or oozing clear fluid;

(p) a reduction in number of days in the preceding week in which thesubject's skin cracked;

(q) a reduction in number of days in the preceding week in which thesubject's skin flaked;

(r) a reduction in number of days in the preceding week in which thesubject experienced dry skin;

(s) an increase in trans epidermal water loss compared to baseline orplacebo control;

(t) an increase in plasma total and free DGLA compared to baseline;

(u) an increase in DGLA:AA ratio compared to baseline or placebocontrol; and/or

(v) a reduction in arterial blood pressure compared to baseline orplacebo control.

In one embodiment, methods of the present invention comprise measuringbaseline levels of one or more markers or parameters set forth in(a)-(v) above prior to dosing the subject or subject group. In anotherembodiment, the methods comprise administering a composition asdisclosed herein to the subject after baseline levels of one or moremarkers or parameters set forth in (a)-(v) are determined, andsubsequently taking an additional measurement of said one or moremarkers.

In another embodiment, upon treatment with a composition of the presentinvention, for example over a period of about 1 to about 12 weeks, about1 to about 8 weeks, or about 1 to about 4 weeks, the subject or subjectgroup exhibits any 2 or more of, any 3 or more of, any 4 or more of, any5 or more of, any 6 or more of, any 7 or more of, any 8 or more of, any9 or more of, any 10 or more of, any 11 or more of, any 12 or more of,any 13 or more of, any 14 or more of, any 15 or more of, any 16 or moreof, any 17 or more of, any 18 or more of, any 19 or more of, any 20 ormore of, any 21 or more of or all 22 of outcomes (a)-(v) describedimmediately above.

In another embodiment, upon treatment with a composition of the presentinvention, the subject or subject group exhibits one or more of thefollowing outcomes:

(a) a reduction in eczema area and severity index (EASI) score relativeto baseline or placebo control of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, at least about 55%, at least about 60%, at leastabout 65%, at least about 70%, at least about 75%, at least about 80%,at least about 85%, at least about 90% or at least about 95%;

(b) a reduction in percentage of area of an anatomical site affected byatopic dermatitis relative to baseline or control of at least about 5%,at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, at least about 55%, at leastabout 60%, at least about 65%, at least about 70%, at least about 75%,at least about 80%, at least about 85%, at least about 90% or at leastabout 95%;

(c) a reduction in investigator's global assessment score relative tobaseline or placebo control of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, at least about 55%, at least about 60%, at least about65%, at least about 70%, at least about 75%, at least about 80%, atleast about 85%, at least about 90% or at least about 95%;

(d) a reduction in intensity of erythema, edema/population,oozing/crusts, excoriation, lichenification and/or dryness relative tobaseline or placebo control of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, at least about 55%, at least about 60%, at least about65%, at least about 70%, at least about 75%, at least about 80%, atleast about 85%, at least about 90% or at least about 95%;

(e) a reduction in erythema, edema/population, oozing/crusts,excoriation, lichenification and/or dryness relative to baseline orplacebo control of at least about 5%, at least about 10%, at least about15%, at least about 20%, at least about 25%, at least about 30%, atleast about 35%, at least about 40%, at least about 45%, at least about50%, at least about 55%, at least about 60%, at least about 65%, atleast about 70%, at least about 75%, at least about 80%, at least about85%, at least about 90% or at least about 95%;

(f) a reduction in body surface area (BSA) affected by atopic dermatitisrelative to baseline or placebo control of at least about 5%, at leastabout 10%, at least about 15%, at least about 20%, at least about 25%,at least about 30%, at least about 35%, at least about 40%, at leastabout 45%, at least about 50%, at least about 55%, at least about 60%,at least about 65%, at least about 70%, at least about 75%, at leastabout 80%, at least about 85%, at least about 90% or at least about 95%;

(g) a reduction in loss of sleep relative to baseline or placebo controlof at least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55%, at least about 60%, at least about 65%, at least about 70%,at least about 75%, at least about 80%, at least about 85%, at leastabout 90% or at least about 95%;

(h) a reduction in occurrence of pruitis (itch) relative to baseline orplacebo control of at least about 5%, at least about 10%, at least about15%, at least about 20%, at least about 25%, at least about 30%, atleast about 35%, at least about 40%, at least about 45%, at least about50%, at least about 55%, at least about 60%, at least about 65%, atleast about 70%, at least about 75%, at least about 80%, at least about85%, at least about 90% or at least about 95%;

(i) a reduction in severity of pruritis as an average of the prior threedays and/or nights on a visual analog scale of at least about 5%, atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, at least about 50%, at least about 55%, at least about60%, at least about 65%, at least about 70%, at least about 75%, atleast about 80%, at least about 85%, at least about 90% or at leastabout 95%;

(j) a reduction in SCORAD score relative to baseline or placebo controlof at least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55%, at least about 60%, at least about 65%, at least about 70%,at least about 75%, at least about 80%, at least about 85%, at leastabout 90% or at least about 95%;

(k) an improved patient-oriented Eczema Measure (POEM) compared tobaseline or placebo control of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, at least about 55%, at least about 60%, at least about65%, at least about 70%, at least about 75%, at least about 80%, atleast about 85%, at least about 90% or at least about 95%;

(l) a reduction in number of days in the preceding week in which thesubject reported that their skin was itchy due to eczema of at leastabout 5%, at least about 10%, at least about 15%, at least about 20%, atleast about 25%, at least about 30%, at least about 35%, at least about40%, at least about 45%, at least about 50%, at least about 55%, atleast about 60%, at least about 65%, at least about 70%, at least about75%, at least about 80%, at least about 85%, at least about 90% or atleast about 95%;

(m) a reduction in number of days in the preceding week in which thesubject reported that their sleep was disturbed due to their eczema ofat least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55%, at least about 60%, at least about 65%, at least about 70%,at least about 75%, at least about 80%, at least about 85%, at leastabout 90% or at least about 95%;

(n) a reduction in number of days in the preceding week in which thesubject experienced skin bleeding of at least about 5%, at least about10%, at least about 15%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, at least about 55%, at least about 60%, atleast about 65%, at least about 70%, at least about 75%, at least about80%, at least about 85%, at least about 90% or at least about 95%;

(o) a reduction in number of days in the preceding week in which thesubject experienced skin weeping or oozing clear fluid of at least about5%, at least about 10%, at least about 15%, at least about 20%, at leastabout 25%, at least about 30%, at least about 35%, at least about 40%,at least about 45%, at least about 50%, at least about 55%, at leastabout 60%, at least about 65%, at least about 70%, at least about 75%,at least about 80%, at least about 85%, at least about 90% or at leastabout 95%;

(p) a reduction in number of days in the preceding week in which thesubject's skin cracked of at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, at least about 55%, at least about 60%, at least about65%, at least about 70%, at least about 75%, at least about 80%, atleast about 85%, at least about 90% or at least about 95%;

(q) a reduction in number of days in the preceding week in which thesubject's skin flaked of at least about 5%, at least about 10%, at leastabout 15%, at least about 20%, at least about 25%, at least about 30%,at least about 35%, at least about 40%, at least about 45%, at leastabout 50%, at least about 55%, at least about 60%, at least about 65%,at least about 70%, at least about 75%, at least about 80%, at leastabout 85%, at least about 90% or at least about 95%;

(r) a reduction in number of days in the preceding week in which thesubject experienced dry skin of at least about 5%, at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 35%, at least about 40%, at least about 45%,at least about 50%, at least about 55%, at least about 60%, at leastabout 65%, at least about 70%, at least about 75%, at least about 80%,at least about 85%, at least about 90% or at least about 95%;

(s) an increase in trans epidermal water loss compared to baseline orplacebo control of at least about 5%, at least about 10%, at least about15%, at least about 20%, at least about 25%, at least about 30%, atleast about 35%, at least about 40%, at least about 45%, at least about50%, at least about 55%, at least about 60%, at least about 65%, atleast about 70%, at least about 75%, at least about 80%, at least about85%, at least about 90% or at least about 95%;

(t) an increase in plasma total and free DGLA compared to baseline of atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, at least about55%, at least about 60%, at least about 65%, at least about 70%, atleast about 75%, at least about 80%, at least about 85%, at least about90% or at least about 95%; and/or

(u) an increase in DGLA:AA ratio compared to baseline or placebo controlof at least about 5%, at least about 10%, at least about 15%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55%, at least about 60%, at least about 65%, at least about 70%,at least about 75%, at least about 80%, at least about 85%, at leastabout 90% or at least about 95%; and/or

(v) a reduction in mean arterial blood pressure of at least about 5%, atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, at least about 50%, at least about 55%, at least about60%, at least about 65%, at least about 70%, at least about 75%, atleast about 80%, at least about 85%, at least about 90% or at leastabout 95%.

In another embodiment, upon treatment with a composition of the presentinvention after a single dose administration or multiple doseadministration, for example over a period of about 1 to about 12 weeks,about 1 to about 8 weeks, or about 1 to about 4 weeks, the subject orsubject group exhibits any 2 or more of, any 3 or more of, any 4 or moreof, any 5 or more of, any 6 or more of, any 7 or more of, any 8 or moreof, any 9 or more of, any 10 or more of, any 11 or more of, any 12 ormore of, any 13 or more of, any 14 or more of, any 15 or more of, any 16or more of, any 17 or more of, any 18 or more of, any 19 or more of, any20 or more of, any 21 or more of or all 22 of outcomes (a)-(v) describedimmediately above.

In another embodiment, upon treatment of a subject or subject group (fedor fasted) with a composition comprising about 200 mg of DGLA to about8000 mg DGLA (administered as one or more dosage units, for example as500 mg or 1 g dosage units equating to total daily DGLA doses of about500 mg, about 1000 mg, about 2000 mg, about 3000 mg, about 4000 mg,about 5000 mg, about 6000 mg, about 7000 mg or about 8000 mg) and aftersingle dose administration or after multiple dose administration, thesubject or subject group exhibits one or more of the following outcomes:

(a) a free DGLA C_(max) (or mean or median C_(max)) of about 400 ng/mlto about 4500 ng/ml, about 500 ng/ml to about 3400 ng/ml, about 600ng/ml to about 3300 ng/ml, about 700 ng/ml to about 3200 ng/ml, forexample about 900 ng/ml, about 1000 ng/ml, about 1100 ng/ml, about 1200ng/ml, about 1300 ng/ml, about 1400 ng/ml, about 1500 ng/ml, about 1600ng/ml, about 1700 ng/ml, about 1800 ng/ml, about 1900 ng/ml, about 2000ng/ml, about 2100 ng/ml, about 2200 ng/ml, about 2300 ng/ml, about 2400ng/ml, about 2500 ng/ml, about 2600 ng/ml, about 2700 ng/ml, about 2800ng/ml, about 2900 ng/ml, about 3000 ng/ml, about 3100 ng/ml, about 3200ng/ml, about 3300 ng/ml, about 3400 ng/ml, about 3500 ng/ml, about 3600ng/ml, about 3700 ng/ml, about 3800 ng/ml, about 3900 ng/ml, about 4000ng/ml, about 4100 ng/ml, about 4200 ng/ml. about 4300 ng/ml about 4400ng/ml or about 4500 ng/ml;

(b) a free DGLA C_(max)/dose (or mean or median C_(max)/dose) of about0.5 (1/kL) to about 3 (1/kL), about 0.6 (1/kL) to about 2.5 (1/kL) orabout 0.7 (1/kL) to about 2 (1/kL), for example about 0.7 (1/kL), about0.8(1/kL), about 0.9 (1/kL), about 1 (1/kL), about 1.5 (1/kL), about 1.6(1/kL), about 1.7 (1/kL) or about 1.8 (1/kL);

(c) a free DGLA AUC₀₋₂₄ (or mean or median AUC₀₋₂₄) of about 1500ng·h/ml to about 12000 ng·h/ml, about 2000 ng·h/ml to about 11000ng·h/ml or about 2500 ng·h/ml to about 10000 ng·h/ml, for example about1000 ng·h/ml, about 1500 ng·h/ml, about 2000 ng·h/ml, about 2500ng·h/ml, about 3000 ng·h/ml, about 3500 ng·h/ml, about 4000 ng·h/ml,about 4500 ng·h/ml, about 5000 ng·h/ml, about 5500 ng·h/ml, about 6000ng·h/ml, about 6500 ng·h/ml, about 7000 ng·h/ml, about 7500 ng·h/ml,about 8000 ng·h/ml, about 8500 ng·h/ml, about 9000 ng·h/ml, about 9500ng·h/ml, about 10000 ng·h/ml, about 10500 ng·h/ml, about 11000 ng·h/ml,about 11500 ng·h/ml or about 12000 ng·h/ml.

(d) a free DGLA AUC₀₋₂₄/dose (or mean or median AUC₀₋₂₄/dose) of about1.5 to about 10 h/kL, about 1.7 to about 8 h/kL or about 2 to about 6h/kL, for example about 2 h/kL, about 2.5 h/kL, about 3 h/kL, about 3.5h/kL, about 4 h/kL, about 4.5 h/kL, about 5 h/kL or about 5.5 h/kL;

(e) a free DGLA t_(max) (h) of about 2 to about 10 hours, about 3 toabout 8 hours, for example about 3 hours, about 4 hours, about 5 hours,about 6 hours, about 7 hours or about 8 hours;

(f) a total DGLA C_(max) (or mean or median total DGLA C_(max)) of about4000 ng/ml to about 45000 ng/ml, about 5000 ng/ml to about 34000 ng/ml,about 6000 ng/ml to about 33000 ng/ml, or about 7000 ng/ml to about32000 ng/ml, for example about 7000 ng/ml, about 7200 ng/ml, about 7500ng/ml, about 8000 ng/ml, about 8500 ng/ml, about 9000 ng/ml, about 9500ng/ml, about 10000 ng/ml, about 11000 ng/ml, about 12000 ng/ml, about13000 ng/ml, about 14000 ng/ml, about 15000 ng/ml, about 16000 ng/ml,about 17000 ng/ml, about 18000 ng/ml, about 19000 ng/ml, about 20000ng/ml, about 21000 ng/ml, about 22000 ng/ml, about 23000 ng/ml, about24000 ng/ml, about 25000 ng/ml, about 26000 ng/ml, about 27000 ng/ml,about 28000 ng/ml, about 29000 ng/ml, about 30000 ng/ml, about 31000ng/ml, about 32000 ng/ml, about 33000 ng/ml, about 34000 ng/ml, or about35000 ng/ml;

(g) a total DGLA C_(max)/dose (or mean or median total DGLAC_(max)/dose) of about 2 (1/kL) to about 25 (1/kL), about 4 (1/kL) toabout 20 (1/kL) or about 5 (1/kL) to about 17 (1/kL), for example about6 (1/kL), about 9 (1/kL), about 14 (1/kL) or about 16 (1/kL);

(h) a total DGLA AUC₀₋₂₄ (or mean or median total DGLA AUC₀₋₂₄) of about15000 ng·h/ml to about 900,000 ng·h/ml, about 20,000 ng·h/ml to about250,000 ng·h/ml or about 25,000 ng·h/ml to about 225,000 ng·h/ml, forexample about 40,000 ng·h/ml, about 210,000 ng·h/ml, about 215,000ng·h/ml or about 435,000 ng·h/ml;

(i) a total DGLA AUC₀₋₂₄/dose (or mean or median total DGLAAUC₀₋₂₄/dose) of about 50 to about 400 h/kL, about 60 to about 250 h/kLor about 70 to about 225 h/kL, for example about 80 h/kL, about 100h/kL, about 110 h/kL or about 215 h/kL;

(j) a total DGLA t_(max) (h) of about 2 to about 25 hours or about 3 toabout 20 hours, for example about 8 hours, about 10 hours, or about 18hours;

(k) a ratio of total DGLA C_(max) to free DGLA C_(max) of about 5:1 toabout 12:1, about 6:1 to about 10:1 or about 7:1 to about 9:1, forexample about 7.7:1, about 8.6:1, about 8.8:1 or about 9.8:1;

(l) a steady state free DGLA plasma level (C_(avg)) or mean or mediansteady state free DGLA plasma level (C_(avg)), after 1 to about 30, 1 toabout 28, 1 to about 14 or 1 to about 10 consecutive days of dailyadministration, of up to about 2000 ng/ml, up to about 750 ng/ml, or upto about 700 ng/ml, for example about 385 ng/ml or about 675 ng/ml;

(m) a steady state total DGLA plasma level (C_(avg)) or mean or mediansteady state total DGLA plasma level (C_(avg)), after 1 to about 30, 1to about 28, 1 to about 14 or 1 to about 10 consecutive days of dailyadministration, of up to 250,000 ng/ml, up to 180,000 ng/ml, up to150,000 ng/ml, up to 125,000 ng/ml or up to 100,000 ng/ml; and/or

(n) a ratio of free DGLA plasma to DGLA skin (e.g. as measured in skinblister fluid) from about 0.2:1 to about 5:1, about 0.5:1 to about 2.5:1or about 0.6:1 to about 1.5:1. In another embodiment, upon treatmentwith a composition of the present invention, for example over a periodof about 1 to about 12 weeks, about 1 to about 8 weeks, or about 1 toabout 4 weeks, the subject or subject group exhibits any 2 or more of,any 3 or more of, any 4 or more of, any 5 or more of, any 6 or more of,any 7 or more of, any 8 or more of, any 9 or more of, any 10 or more of,any 11 or more of, any 12 or more of, any 13 or more of, any 14 or moreof, any 15 or more of, any 16 or more of, any 17 or more of, any 18 ormore of, any 19 or more of, any 20 or more of, any 21 or more of or all22 of outcomes (a)-(n) described immediately above.

In another embodiment, upon treatment of fasted and fed subjects orfasted and fed subject groups with a composition comprising about 200 mgof DGLA to about 8000 mg DGLA (administered as one or more dosage units,for example as 500 mg or 1 g dosage units equating to total daily DGLAdoses of about 500 mg, about 1000 mg, about 2000 mg, about 3000 mg,about 4000 mg, about 5000 mg, about 6000 mg, about 7000 mg or about 8000mg) and after single dose administration or after multiple doseadministration, the subject or subject group exhibits one or more of thefollowing outcomes:

(a) a ratio of free DGLA C_(max) fasted:fed between about 1:1 to about5:1, for example of about 2.5:1, of about 3:1 or of about 3.5:1;

(b) a ratio of free DGLA AUC₀₋₂₄ fasted:fed between about 1:1 and about5:1, for example of about 1.5:1, of about 2:1, or of about 2.5:1;

(c) a ratio of total DGLA C_(max) fasted:fed between about 1:1 to about5:1, for example of about 1:1, of about 1.5:1 or of about 2:1; and/or

(d) a ratio of total DGLA AUC₀₋₂₄ fasted:fed between about 1:1 and about5:1, for example of about 1.5:1, of about 2:1 or of about 2.5:1.

In one embodiment, a DGLA-containing composition of the inventioncomprises the following fatty acid fingerprint:

C18:1n-9 <LOD-<5% C18:2n-6 <LOD-<5% 20:3ω6-DGLA NLT 95 isomerA <LOD-<5%C20:4n-6 + isomerB <LOD-<5% Total unidentified NMT 2 related substances

In one embodiment, a DGLA-containing composition of the inventioncomprises the following fatty acid fingerprint:

Fatty Acid Profile (Area % FAMEs by GC) 20:3ω6-DGLA NLT 95 RelatedSubstances 20:2ω6 <LOD-<5% 20:3ω3 <LOD-<5% 20:4ω6 <LOD-<5% 20:4ω3<LOD-<5% 20:5ω3 <LOD-<5% Total unidentified related substances NMT 2

An illustrative DGLA-containing composition of the invention comprisesthe following fatty acid fingerprint:

C16:0 <LOD-<5% C18:1n-7 <LOD-<5% C18:1n-9 <LOD-<5% C18:2n-6 <LOD-<5%C18:3n-6 <LOD-<5% C20:3n-3 <LOD-<5% 20:3n-6-DGLA NLT 95 C20:4n-6<LOD-<5% Total unidentified related NMT 2 substances

In one embodiment, a DGLA-containing composition of the inventioncomprises the following fatty acid fingerprint:

Fatty Acid Profile (Area % FAMEs by GC) 20:3ω6-DGLA NLT 95 RelatedSubstances 16:0 <LOD-<5% 18:3n-6 alcohol methyl ether <LOD-<5% 18:3n-6alcohol formate <LOD-<5% 16:3n-3 <LOD-<5% 18:1n-9 <LOD-<5% 18:1n-7<LOD-<5% 19:3 <LOD-<5% 20:1n-9 <LOD-<5% 20:2n-6 <LOD-<5% 20:2n-3 + DGLAisomer <LOD-<5% 20:3n-3 <LOD-<5% 20:4n-3 <LOD-<5% Methyl7,11,14-eicosatrienoate (DGLA isomer) <LOD-<5% 22:5n-3 <LOD-<5% Totalunidentified related substances NMT 2

In one embodiment, a DGLA-containing composition of the inventioncomprises the following fatty acid fingerprint:

C20:3 n-6 (DGLA - triglycerides) Min. 30% C16:0 Max. 26% C18:0 Max. 12%C18:1 n-9 Max. 15% C18:2 n-6 Max. 15% C18:3 n-6 Max. 5% C20;4 n-6 Max.1% C22:0 Max. 5% C24:0 Max. 15%

EXAMPLES Example 1

Three batches of pharmaceutical compositions comprising DGLA (with 2000pm dl-alpha tocopherol) filled into gelatin capsules were prepared asshown in Table 1.

TABLE 1 Batch Number DGLA (mg/Capsule) Gelatin Description E09726/1 250Standard acid bovine gelatin E09726/2 250 Lime bone gelatin with a lowermolecular weight (Mw) E09727 500 Standard acid bovine gelatin

The capsules shells included the following excipients: gelatin, purifiedwater, glycerol, titanium dioxide, and the processing aids lecithin andmedium chain triglyceride.

Additional batches of DGLA capsules were also prepared including DGLAFFA (stabilized with a nominal 2000 ppm dl-alpha tocopherol) in capsulescontaining gelatin, polysorb or mixture of gylycerol/polysorb, purifiedwater, titanium dioxide, and the processing aids lecithin andmedium-chain triglyceride as shown in Table 2.

TABLE 2 Batch DGLA Number (mg/Capsule) Gelatin Description PlasticizerE09778 500 Lime bone gelatin with Glycerol a lower molecular weight (Mw)E09777/01 500 Lime bone gelatin with Polysorb (D-sorbitol a lowermolecular and 1,4-sorbitan weight (Mw) sugar alcohols in water solution)E09777/02 500 Lime bone gelatin with Gycerol + Polysorb a lowermolecular weight (Mw) E09777/03 500 Lime bone gelatin with Polysorb alower molecular weight (Mw) - Even Lower Mw (Advanced RXL Gelatine)

Capsule shell compositions for each of the batches are shown below inTables 3 and 4.

TABLE 3 Unit Quantity mg/capsule Active Substance 500 mg DGLA E09778E09777/01 E09777/02 E09777/03 Wet Gelatin Shell Mass Gelatin 128.97128.97 128.97 128.97 (RXL) (RXL) (RXL) (RXL Adv.)¹ Total Glycerol  67.702.94 35.32 2.94 Polysorb N/A 64.76  32.38 64.76  Purified Water² 100.38100.38  100.38  100.38  Titanium Dioxide  2.94 2.94  2.94 2.94 LecithinTrace Trace Trace Trace Triglycerides Medium Trace Trace Trace TraceChain ¹RXL gelatin contains a lower number of high molecular weightpolymers (~5% >200,000 Da)

TABLE 4 Unit Quantity E09726/01 mg/capsule % w/w mg/capsule % w/w ActiveSubstance DGLA 500 100 250 100 Wet Gelatin Shell Mass² Gelatin (not RXL)132.35 44.12 87.79 44.12 Total Glycerol 76.76 25.59 50.92 25.59 PurifiedWater 87.95 29.31 58.33 29.31 Glycerol and — — — — Polysorb? TitaniumDioxide 2.94 0.98 1.95 0.98 Lecithin Trace Trace Trace TraceTriglycerides Trace Trace Trace Trace Medium Chain

Stability testing of the above capsules was performed. Capsules fromeach batch were maintained for up to 6 months and assessed using aqualitative or quantitative USP 2040 Disintegration and Dissolution testprotocol. Results are shown in Tables 5-7.

TABLE 5 Stability Data for DGLA Softgel capsules: Qualitative RuptureTest Results Specification: read and record (min) Results (Months): 1 36 40° 2 25° C./ 30° C./ 40° C./ 25° C./ 30° C./ 40° C./ C./ 40° C./ 60%RH 65% RH 75% RH 60% RH 65% RH 75% RH 0 75% 75% Without With WithoutWith Without With Without With Without With Without With Batches N/A RHRH pepsin pepsin pepsin pepsin pepsin pepsin pepsin pepsin pepsin pepsinpepsin pepsin E09726/ 7 6 >30/22⁽¹⁾ 8 N/A 1 NP >30 14 >30 >30 >30 2 01caps caps on on 6 6 >30 >30 E09726/ 4 4 4 3 N/A 1 6 1 10 2 9 12 15 02caps caps caps on 6 on 6 on 6 >30 >30 >30 E09727 >30 4 caps 10 >30NP >30 15 >30 14 >30 16 >30 3 on 6 caps >30 on 6 >30

TABLE 6 DGLA glyceride percentages Batch Time 25 30 40 Time point(months) DGLA glycerides % E09727 0 0 0 0 1 0 0 0.54 2 0 0 1.19 3 0.410.67 2.1 6 0.53 1.43 5.81 E09726/1 0 0 0 0 1 0 0 0.56 2 0 0 1.35 3 0.370.58 2.31 6 0.68 1.42 6.8 E09726/2 0 0 ND 0 1 0 ND ND 2 0 ND 1.63 3 0.51ND ND 6 1.16 ND 8.55

TABLE 7 Mono, Di and Triglycerides DGLA Quantitative Storage Acid assay(% DGLA) esters (%) rupture test Conditions value Read and Read and Readand record Specifications 40° C./75% RH 176-184 record record (mean %)Tier I E09778 (RXL + 1 month 178 0.98 1.8 to 2.6² 15 min 94 Glycerol) 30min 102 45 min 102 60 min 102 Tier II stage 2 3 months 172 3.08 N/A 15min N/A 30 min 92 45 min 96 60 min 97 Tier I E09777/01 (RXL + 1 month180 N/A 1.8 to 2.9² 15 min 81 Polysorb) 30 min 91 45 min 96 60 min 102Tier II 3 months 182 N/A Not available 15 min N/A 30 min 97 45 min 99 60min 98 Tier I E09777/02 (RXL + 1 month 177 <0.5 0.0 to 3.4² 15 min 95Glycerol + 30 min 101 Polysorb) 45 min 101 60 min 102 Tier I 3 months177 3.1 Not avalable 15 min 92 30 min 97 45 min 97 60 min 98 Tier IE09777/03 (RXL adv. + 1 month  171⁽²⁾ N/A 2.7 to 3.6² 15 min 88Polysorb) 30 min 97 45 min 99 60 min 99 Tier I 3 months 174 N/A Not 15min 87 Available 30 min 94 45 min 95 60 min 95

As seen above, there was a slowdown in dissolution rate in water overtime for capsules formulated with glycerol and standard acid bovinegelatin (E09726/01, and E09727). There was a DGLA release rate ofgreater than 30 mins after 6 months at 40° C./75% RH in simulatedgastric fluid (pH 1.2, pepsin).

A DGLA release rate of less than 30 mins after 6 months 40° C./75% RHwas only achieved in simulated gastric fluid (pH 1.2, pepsin) withcapsules containing lime bone gelatin with a lower molecular weight (Mw)(E09777102).

There was a significant increase in DGLA glyceride formation over timein DGLA capsule shells containing glycerol (Table 4). This wastemperature dependent with highest concentrations of DGLA formed at 40°C. 75% RH.

Polysorb is commonly used as a hydrophilic plasticizer to limit exchangebetween capsule fill media and shell. D-Sorbitol and 1,4-sorbitan have ahigher MW than glycerol which limits its mobility through the gelatinshell. Despite this, there was still interaction of D-Sorbitol and1,4-sorbitan to form DGLA FFA esters in batches E09777 1/2 and 3.

There was no reduction in acid value of the DGLA for batches formulatedwith D-Sorbitol and 1,4-sorbitan (E09777 1/02/3) whereas there was areduction in acid value for E09778 formulated with glycerol.

There was no slowdown in dissolution rate in water over time forcapsules formulated with D-Sorbitol and 1,4-sorbitan (E09777103). TheDGLA release rate was less than 30 minutes in water after 3 months 40°C./75% RH.

Example 2

A randomised, double-blind, placebo-controlled, phase II study to assessthe efficacy and safety of orally administered DGLA to patients withmoderate to severe atopic dermatitis will be conducted. Approximately100 male or female subjects, aged 18 years or older, with moderate tosevere atopic dermatitis (AD) will be included in this study.

Atopic dermatitis is a chronic inflammatory skin disorder characterizedby the presence of pruritus, papules, lichenification, excoriations,xerosis and oozing. The prevalence of AD has increased in the last fewdecades, affecting up to 20 percent of young children with the majorityof cases starting in children younger than 5 years of age. Most casesimprove by adulthood.

AD is a multifactorial disease, with genetics, environment, and impairedimmune response being the predominant factors. Dendritic cells, Tlymphocytes, macrophages, keratinocytes, mast cells, and eosinophils allplay a role in AD by releasing proinflammatory cytokines and chemokineswhich induce the inflammatory response characteristic of atopicdermatitis lesions.

Most of currently approved treatments for AD are topical and includecorticosteroids, pimecrolimus and tacrolimus. Corticosteroids have beenthe mainstay of treatment for AD and different potencies andformulations are available. Topical corticosteroids are effective forthe treatment of AD but their use is limited by the potential for localside effects such as skin atrophy and striae. Systemic absorption ofcorticosteroids can also induce diabetes, cataracts, osteoporosis andsuppression of the hypothalamic pituitary axis. In addition, transientimprovement is often followed by a rebound flare-up on discontinuationof treatment. Other treatments available include topical calcineurininhibitors (e.g., pimecrolimus, tacrolimus), and coal tar preparations.Cases of lymphoma have been reported in patients treated withcalcineurin inhibitors. Patients with more severe disease are treatedwith ultraviolet B and A phototherapy or oral agents such ascorticosteroids, cyclosporine, mycophenolate mofetil, methotrexate andazathioprine.

Dihomo-gamma-linolenic acid (DGLA) is an essential fatty acid foundnaturally in the body as the 2 carbon elongation product ofgamma-linolenic acid (GLA). DGLA is metabolized by cyclooxygenase (COX)and lipoxygenase (LPO) enzymes to form eicosanoids, prostaglandins (PG)of the 1 series and also hydroxyl-fatty acid as 15 hydroxyeicosatrienoicacid DGLA (15 HETrE).

Study Hypothesis and Objectives

DS107G is superior to placebo for improvement of EASI score in patientswith moderate to severe atopic dermatitis.

Primary objective: To compare the efficacy of two doses of orallyadministered DS107G capsules versus placebo, in the treatment of adultpatients with moderate to severe atopic dermatitis.

Secondary objective: To assess the safety of two doses of orallyadministered DS107G capsules versus placebo, in adult patients withmoderate to severe atopic dermatitis.

Study Endpoints Primary Endpoint

Proportion of patients achieving an IGA (Investigator Global Assessment)of 0 (clear) or 1 (almost clear) and a decrease of at least 2 points inIGA at Week 8.

Secondary Endpoints

Change from baseline in IGA at week 2, 4 and 8;

Change from baseline in EASI (Eczema Area and Severity Index) at week 2,4 and 8;

Proportion of patients achieving at least a 1-point decrease in IGA atweek 8;

Change from baseline in the Patient Orientated Eczema Measure (POEM) atweek 2, 4 and 8;

Change from baseline in the Dermatology Life Quality Index (DLQI) scoreat week 2, 4 and 8;

Change from baseline in SCORAD at week 2, 4 and 8;

Change from baseline in the patient's Visual Analog Scale (VAS) pruritusscore at Week 2, 4 and 8;

Change from baseline in Body Surface Area (BSA) at Week 2, 4 and 8;

Number of treatment-emergent adverse events (TEAEs) in each treatmentgroup;

Exploratory Endpoints

Change from baseline in Trans epidermal water loss (TEWL) at week 2, 4and 8 (selected sites only);

Plasma total and free DGLA concentrations at Baseline, weeks 4 and 8;

Plasma total fatty acid profile at Baseline, week 4 and week 8 (sampleto be retained and analyzed at a later date);

Study Design

Approximately 100 patients with moderate to severe atopic dermatitiswill be included in this multicenter, double blind, placebo controlled,phase Ila study. All subjects will sign an informed consent and undergoscreening for study eligibility. Subjects will be randomized (1:1) atbaseline visit to either receive oral 2 g DS107G (DGLA capsules providedas opaque, oval soft gelatin capsules containing 500 mg DGLA), 1 gDS107G or placebo once daily for 8 weeks in a fasting state. Enrollmentwill be stratified by disease severity (based on IGA) and a maximum of30% of patients with mild atopic dermatitis (as defined by an IGA scoreof 2) will be recruited.

Subjects will come to the clinic on 6 occasions: at screening, baseline,week 2, week 4, week 8 (end of treatment/early termination) and week 10(follow-up). All subjects will exit the study at the Week 10 visit. Theprimary efficacy variable will be the proportion of patients achievingan IGA of 0 (clear) or 1 (almost clear) and a decrease of at least 2points in IGA at week 8. Secondary efficacy variables will include IGAat other visits, pruritus (obtained from the SCORAD visual analogscale), EASI, BSA, POEM, DLQI, SCORAD and TEWL (for selected sitesonly). Safety will be assessed through adverse events, physicalexamination, vital signs and safety laboratory tests (includingpregnancy tests for women of childbearing potential). Pharmacokineticsamples will be obtained at Baseline (Day 0), week 4 and week 8 visitsin order to measure total and free DGLA plasma trough levels. Separateplasma samples will be retained for later analysis of total fatty acidprofile and interleukin profile.

Study Population

Approximately 100 subjects with moderate to severe atopic dermatitis asper IGA score and a BSA of minimum 10% may be included in this study.Subjects will be men or women, 18 years or older.

Inclusion Criteria:

Male or female subject aged 18 years and older on the day of signing theinformed consent form (ICF).

Clinically confirmed diagnosis of active atopic dermatitis according toHanifin and Rajka criteria (Appendix G).

Moderate to severe atopic dermatitis at baseline as defined by an IGA ofminimum 3 at baseline visit.

Atopic dermatitis covering minimum 10% of the body surface area atbaseline.

Body mass index (BMI) is between 18 and 35 kg/m2 inclusively.

Female patients of childbearing potential must use adequatecontraception or have a sterilized partner for the duration of thestudy: systemic hormonal contraceptives, intrauterine device or barriermethod of contraception in conjunction with spermicide, or agree tosexual abstinence. Hormonal contraceptives must be on a stable dose forat least one month before baseline. Note: Women of non-child bearingpotential are:

-   -   women who have had surgical sterilization (hysterectomy or        bilateral oophorectomy or tubal ligation);    -   women greater than 60 years of age;    -   women greater than 40 and less than 60 years of age who have had        a cessation of menses for at least 12 months and a        follicle-stimulating hormone (FSH)        test confirming non-childbearing potential (FSH≧40 mlU/mL) or        cessation of menses for at least 24 months without FSH levels        confirmed.

Patients who are able and willing to stop treatment for atopicdermatitis throughout the study (except for allowed emollients);

Capable and willing to give signed informed consent and the consent mustbe obtained prior to any study related procedures.

Exclusion Criteria

Female patients with positive pregnancy test at screening or Day 0 visit(baseline) or lactating women.

Any clinically significant controlled or uncontrolled medical conditionor laboratory abnormality that would, in the opinion of theinvestigator, put the patient at undue risk or interfere withinterpretation of study results.

Clinically significant impairment of renal or hepatic function.

Other skin conditions that might interfere with atopic dermatitisdiagnosis and/or evaluation (such as psoriasis or current viral,bacterial and fungal skin infections).

History of hypersensitivity to any substance in DS107G or placebocapsules.

Use of biologics 3 months prior to start of treatment/Day 0 visit(baseline), or 5 half-lives (whichever is longer).

Use of systemic treatments (other than biologics) that could affectatopic dermatitis less than 4 weeks prior to baseline visit (Day 0),e.g. retinoids, calcineurin inhibitors, methotrexate, cyclosporine,hydroxycarbamide (hydroxyurea), azathioprine and oral/injectablecorticosteroids; Intranasal corticosteroids and inhaled corticosteroidsfor stable medical conditions are allowed.

Treatment with any experimental drug within 30 days prior to Day 0 visit(baseline), or 5 half-lives (whichever is longer).

Excessive sun exposure, use of tanning booths or other ultraviolet (UV)light sources 4 weeks prior to Day 0 visit (baseline) and/or is planninga trip to sunny climate or to use tanning booths or other UV sourcesbetween screening and follow-up visits.

Use of any topical medicated treatment for atopic dermatitis 2 weeksprior to start of treatment/Day 0 visit (baseline), including but notlimited to, topical corticosteroids, calcineurin inhibitors, tars,bleach, antimicrobials and bleach baths.

Use of topical products containing urea, ceramides or hyaluronic acid 2weeks prior to Day 0.

Use of anti-histamines for atopic dermatitis within 2 weeks of baseline.

Significant uncontrolled cardiovascular (a history of ECG abnormalitiesthat are clinically significant in the opinion of the investigator),neurologic, malignant, psychiatric, respiratory or hypertensive disease,as well as diabetes and arthritis.

Medical history of chronic infectious disease (e.g., hepatitis B,hepatitis C or infection with human immunodeficiency virus).

History of clinically significant drug or alcohol abuse in the last yearprior to Day 0 (baseline).

Study Restrictions

Each subject will be questioned on the specific points listed belowprior to drug administration. If a subject admits a non-compliance withthese restrictions, the Principal Investigator (or designee) and/or thesponsor will decide whether or not the subject will be permitted toremain in the study. Non-compliance with these restrictions will berecorded.

Subjects will be instructed to abstain from planning a trip to sunnyclimate or use of tanning equipment between screening and follow-upvisits.

Subjects will be instructed to abstain from using any drugs/treatmentsthat may influence atopic dermatitis (refer to exclusion criteria andprohibited therapies or procedures section) throughout the study.

Subjects will be required to start fasting at least 8 hours before drugadministration upon waking. Fasting will continue for at least 60minutes following drug administration, after which subject can havebreakfast. Water will be allowed at all times during the fasting period,but no other fluids will be permitted. Medication(s) for otherconditions that are permitted in the study can be taken as usual.

For Baseline (Day 0), week 4 and week 8 visits, a blood draw will beperformed for PK analysis. PK samples must be taken pre-dose; therefore,study drug administration will occur during the visit for Day 0 and week4 visits. Because dosing will occur at the clinic on Day 0 and week 4,subjects will be required to fast for at least 8 hours prior to studydrug administration and will be allowed to have a meal 60 minutes afterstudy drug administration.

Discontinuations

Subjects have the right to withdraw from the study at any time for anyreason without penalty. The investigator also has the right to withdrawsubjects from the study if he feels it is in the best interest of thesubject or if the subject is uncooperative or non-compliant. It isunderstood by all concerned that an excessive rate of withdrawal canrender the study un-interpretable; therefore, unnecessary withdrawal ofsubjects should be avoided. Should a subject decide to withdraw, allefforts will be made to complete and report the observations,particularly the follow-up examination, as thoroughly as possible.

The investigator or one of his or her staff members should contact thesubject either telephone or through a personal visit to determine ascompletely as possible the reason for the withdrawal, and record thereason in subject's source document and CRF. A complete final earlytermination (week 8) evaluation at the time of the subject's withdrawalshould be made with an explanation of why the subject is withdrawingfrom the study. If the reason for removal of a subject is an adverseevent or an abnormal laboratory test result, the principal specificevent or test will be recorded. Subjects who discontinue the studybefore week 8 visit will be asked, if they agree, to come for a lastassessment (early termination visit).

Reasons for Discontinuation Include:

The investigator decides that the subject should be withdrawn. If thisdecision is made because of a serious or persistent adverse event,laboratory abnormality, or intercurrent illness, the study drug is to bediscontinued and appropriate measures are to be taken. The investigatorwill notify the Sponsor or designee immediately.

The subject or attending physician requests that the subject bewithdrawn from the study.

The subject for any reason requires treatment with another therapeuticagent that has been demonstrated to be effective for treatment of thestudy indication. In this case, discontinuation from the study occursimmediately upon introduction of the new agent.

The subject is lost to follow-up, in this case, a reasonable attempt tocontact the subject and ascertain his/her status must be made and theseattempts must be documented.

Serious protocol violation, including persistent non-compliance.

The Sponsor or Regulatory Authorities, for any reason, stops the study.All subjects will be discontinued from the study and notified of thereasons for the discontinuation.

Pregnancy at any time during the study.

Other: the subject may withdraw from the study for any other reason,including withdrawal of consent.

Treatment Treatment Administration

Subjects who fulfill all the inclusion and none of the exclusioncriteria may be accepted in the study. Each subject must read and signan informed consent form prior to any screening procedures beingperformed. This study involves a comparison of DS107G (2 g) withplacebo, administered orally once daily upon waking for a total durationof 8 weeks. The last study drug administration should occur on the daypreceding week 8 visit/Early Termination (ET) visit. Subjects will berandomized to one of the two treatment groups in a 1:1 ratio:

Treatment group A: 2 grams DS107G (4 capsules).

Treatment group B: 2 grams placebo capsules (4 capsules).

Subjects will be required to start fasting at least 8 hours before drugadministration upon waking. Fasting will continue for at least 60minutes following drug administration, after which subjects can havebreakfast. Water will be allowed at all times during the fasting period,but no other fluids will be permitted. Medication(s) for otherconditions that are permitted in the study can be taken as usual.

Blister packs will consist of 7 rows of 4 capsules. Each row constitutesone daily dose. Subjects will be instructed to take the 4 capsules fromleft to right, top to bottom.

Study Treatment

DS107G capsules will be provided as opaque, oval soft gelatin capsulescontaining 500 mg of DGLA free fatty acid (FFA).

Placebo capsules will be also provided as opaque, oval soft gelatincapsules containing 500 mg of liquid paraffin.

DS107G capsules will be supplied in manufactured form (blinded),packaged in aluminum foil blisters of 28 units. Placebo will bepresented in identical blisters and packs and stored/packaged the sameas DS107G capsules. Study medication will be labelled according to USand Canadian regulations.

The study medication will be provided by the sponsor to the investigatorand will be kept, on site, in a locked room or cabinet with limitedaccess. DS107G and placebo capsules should be stored at a controlledroom temperature between 15-30° C. and will only be supplied to subjectsin the trial under the supervision of the investigator.

Study drug will be dispensed by the study site to the subject at eachstudy visit. Subjects are to return all study drug blister packs (usedand unused) to the study site. The capsules within blister packs will becounted prior to dispensing and upon return and the counts will berecorded in the source documents and eCRF. Each subject is to beinstructed on the importance of returning study drug at the next studyvisit. If a subject does not return study drug, he/she will beinstructed to return it as soon as possible.

The investigator is responsible for maintaining accurate records of thestudy medication received initially, the study drug dispensed/used, thereturned medication by subjects and the medication destroyed or returnedto the Sponsor or designee. All study drug accountability forms andtreatment logs must be retained in the Investigator's study file. Theserecords must be available for inspection by the Sponsor, its designeesor by regulatory agencies at any time.

Used drug boxes/blister packs will be stored safely until destructionand must be accounted for by the investigator. The study monitor willperform drug accountability for all study drug at the site and assist inreturning study drug, including used and unused study drug to theSponsor or designee. After verification of the drug accountability bythe sponsor, the investigator will ensure proper destruction or returnof the remaining study product.

Any study medication accidentally or deliberately destroyed will beaccounted for. Any discrepancies between amounts dispensed and returnedwill be explained.

Drug inventory and accountability records will be maintained at eachsite as per GCP/ICH guidelines. Approximately 100 patients will berandomized into double-blind treatment groups in a 1:1 ratio by anInteractive Web Response System (IWRS) or Interactive Voice ResponseSystem (IVRS), as follows:

Treatment group A: 2 grams DS107G (4 capsules).

Treatment group B: 2 grams placebo capsules (4 capsules).

A randomization list permuted blocks and stratified by site will begenerated by Dignity Sciences or its designee. The randomizationschedule with study drug assignments will be generated prior to thestart of the study and will be known only to the individuals responsiblefor labeling the study drug. The IVRS or IWRS will assign a study drugkit number to each subject and the contents will be based on therandomization code.

At the investigational site, each subject will be assigned a patientidentifier number during screening that will be used on all patientdocumentation. The patient identifier number will contain the sitenumber and the patient number assigned in numerical order at thescreening visit (e.g.: 02-010 for the tenth patient screened at the site#02). Numbers will be assigned in ascending order starting with 001.

Rationale for Selection and Timing of Doses in the Study

Doses up to 4 g have been well tolerated in healthy subjects. The doseof 2 g has been selected as the tested dose in the current study basedprimarily on the pharmacokinetic results from the Phase I trial whichsuggested saturable skin levels of total DGLA with repeated oral dosingof greater than 2 g per day. In addition the following factors wereconsidered:

there were less frequent transient gastrointestinal instances recordedat a dose of 2 g compared to 4 g daily.

the number of capsules (4) administered daily. A higher dose would bepossible but is deemed less desirable as too many capsules may have anegative impact on patient adherence to treatment.

Breaking of Study Blinding

At all times, treatment and randomization information will be keptconfidential and will not be released to the investigator, the studystaff, the CRO or the sponsor's study team until following theconclusion of the study.

Blinding codes should only be broken in emergency situations for reasonsof subject safety. The method will be either a manual or electronicprocess. When the blind for a subject has been broken, the reason mustbe fully documented. Whenever possible, the investigator should contactthe Sponsor or its designee before breaking the blind. If the blind isbroken, the investigator should promptly inform the Medical Monitor.Documentation of breaking the blind should be recorded with the date andtime the blind was broken, and the names of the personnel involved.

The subject for whom the blind has been broken will be discontinued fromthe study and undergo the early termination (ET) procedures (Week 8visit). In cases where there are ethical reasons to have the subjectremain in the study, the investigator must obtain specific approval fromthe Sponsor or its designee for the subject to continue in the study.

Concomitant Therapy

All medications (including over-the-counter drugs, vitamins, andantacids) taken ≦4 weeks prior to screening and throughout the studymust be recorded. All medications taken for atopic dermatitis in the 2months prior to screening must be recorded.

Medication entries should be specific to the generic name. Trade namemay be used for combination drugs. Entries will include the dose, unit,and frequency of administration, route of administration, start date,discontinuation date, and indication. If the medication is discontinued,or the dosage changed, these details must be recorded.

The Investigator should assess any concomitant procedures, medications,and dietary supplements for acceptability that are not explicitlyprohibited.

Permitted Therapies

Emollients

Subjects can apply a bland emollient of their choice on their skin,including AD lesions, provided that emollient use is initiated at least2 weeks prior to Day 0 and continues at the same frequency and on thesame skin areas throughout the study. Subjects will be requested toavoid using emollients containing any of the following ingredients:

(1) Urea

(2) Ceramide

(3) Hyaluronic acid

Every effort should be made to keep the same emollient throughout thestudy. The commercial name of the selected emollient(s) will be recordedin the source document and the eCRF. No other products may be applied tothe lesions during the study.

Other Permitted Therapies

Non-sedative anti-histamines (e.g. loratadine, fexofenadine) are allowedduring the study only if used to treat medical conditions other thanatopic dermatitis. Such medications are allowed during the study only ifthe subject has been on a stable dose for at least 2 weeks prior to Day0 and continues to use the same agent everyday throughout the study.

Inhaled and intranasal corticosteroids for stable medical conditions areallowed.

Prohibited Therapies or Procedures

The following topical therapies or procedures are prohibited during thestudy for all subjects:

Topical medicated treatments that could affect atopic dermatitis,including but not limited to:

topical corticosteroids

calcineurin inhibitors

tars

bleach

antimicrobials

bleach baths

Any topical product containing urea, ceramides or hyaluronic acid

Systemic therapy that could affect atopic dermatitis, e.g. retinoids,calcineurin inhibitors, methotrexate, cyclosporine, hydroxycarbamide(hydroxyurea), azathioprine and oral/injectable corticosteroids

Anti-histamines (except non-sedative anti-histamine)

Any biological agent

UVA or UVB phototherapy

Psoralen+Ultraviolet A (PUVA) therapy

Excessive sun exposure or use of tanning booth

Any investigational agent

Assessment of Compliance

Treatment compliance will be assessed at each visit by directquestioning, review of the subject's compliance log and capsule count,and will be based on the latter. Subjects will be given a paper diary ateach visit along with study medication. Subjects will indicate anymissed doses on the diary, as well as the timing of the last foodingestion prior to study drug administration and food ingestionfollowing study drug administration. Subjects will be instructed tobring all capsules and blister packs (used and unused) and compliancelog to the next study visit. Any deviation from the prescribed dosageregimen will be recorded in the source document and in the eCRF.Subjects who are significantly noncompliant will be counseled.

Study Procedures

Please refer to Appendix A for a flowchart of procedures to perform ateach visit.

Screening, Visit 1 (Day −30 to −1)

Screening evaluation will only be performed after the subject has agreedto participate and has signed and dated the informed consent form. Notreatment or trial related procedures will be initiated before theinformed consent is signed. Day 0 visit must be performed, at thelatest, 30 days after the screening visit.

Screening evaluation will be performed according to inclusion andexclusion criteria. If the subject fulfils all inclusion criteria and noexclusion criteria, the subject may be included in the study.

The following procedures will be performed at the screening visit:

Informed consent;

Review of Inclusion-Exclusion criteria, including review of Hanifin andRajka criteria (Appendix G);

Assign subject identifier number (Site number-Subject number);

Demographics;

Concomitant medications;

Medical/surgical history;

Physical examination;

Vital signs;

BMI;

Safety labs (chemistry, coagulation, hematology and urinalysis);

Serum pregnancy test (women of childbearing potential only) and FSHlevel test for women greater than 40 and less than 60 years of age whohave had a cessation of menses for at least 12 months but less than 24months;

BSA evaluation;

IGA;

Baseline, Visit 2 (Day 0)

Subjects are required to fast for at least 8 hours prior to study drugadministration. They will be allowed to have a meal 60 minutes afterstudy drug administration.

The following procedures will be performed at this visit:

(1) Confirm eligibility with inclusion and exclusion criteria

(2) Update or confirm medical/surgical history

(3) Concomitant medications

(4) Vital signs

(5) BMI

(6) Safety labs and biomarkers of inflammation (chemistry, coagulation,hematology, urinalysis, and interleukin profile).

(7) Urine pregnancy test (women of childbearing potential only)

(8) Pre-dose blood draw pharmacokinetics

(9) Blood draw total fatty acid profile

(10) BSA evaluation

(11) IGA

(12) EASI assessment

(13) SCORAD assessment (including VAS pruritus assessment)

(14) POEM questionnaire

(15) DLQI questionnaire

(16) TEWL assessment (for selected sites only)

(17) Randomize subject in IVRS/IWRS

(18) Study drug administration

(19) Dispensing of study drug

(20) Dispensing of Subject Compliance Log

(21) Adverse events evaluation (after first study drug administration)

Week 2, Visit 3 (Day 14±2 days)

The following procedures will be performed at this visit:

(1) Vital signs

(2) Urine pregnancy test (women of childbearing potential only)

(3) BSA evaluation

(4) IGA

(5) EASI assessment

(6) SCORAD assessment (including VAS pruritus assessment)

(7) POEM questionnaire

(8) DLQI questionnaire

(9) TEWL assessment (for selected sites only)

(10) Collection and dispensing of study drug Review, collection anddispensing of Subject Compliance log

(11) Capsule count

(12) Concomitant medications

(13) Adverse events evaluation

Week 4, Visit 4 (Day 28±2 days).

Subjects are required to fast for at least 8 hours prior to study drugadministration. They will be allowed to have a meal 60 minutes afterstudy drug administration.

The following procedures will be performed at this visit:

(1) Vital signs

(2) Urine pregnancy test (women of childbearing potential only)

(3) Physical examination

(4) Safety labs (chemistry, coagulation, hematology, urinalysis andinterleukin profile)

(5) Pre-dose blood draw pharmacokinetics (if study medication was takenprior to the visit, subject must come back the next day)

(6) Blood draw total fatty acid profile

(7) BSA evaluation

(8) IGA

(9) EASI assessment

(10) SCORAD assessment (including VAS pruritus assessment)

(11) POEM questionnaire

(12) DLQI questionnaire

(13) TEWL assessment (for selected sites only)

(14) Study drug administration (instruct subject that last dose shouldoccur on the day preceding week 8 visit)

(15) Collection and dispensing of study drug

(16) Review, collection and dispensing of Subject Compliance log

(17) Capsule count

(18) Concomitant medications

(19) Adverse events evaluation.

Week 8, Visit 5 (Day 56±2 days) (End of Treatment/Early TerminationVisit).

The following procedures will be performed at this visit:

(1) Ongoing medical history review

(2) Vital signs

(3) Physical examination

(4) BMI

(5) Safety labs (chemistry, coagulation, hematology and urinalysis)

(6) Serum pregnancy test (women of childbearing potential only)

(7) Blood draw pharmacokinetics (if study medication was taken prior tothe visit, subject must come back the next day)

(8) Blood draw total fatty acid profile

(9) BSA evaluation

(10) IGA

(11) EASI assessment

(12) SCORAD assessment (including VAS pruritus assessment)

(13) POEM questionnaire

(14) DLQI questionnaire

(15) TEWL assessment (for selected sites only)

(16) Collection of study drug

(17) Review and collection of Subject Compliance log

(18) Capsule count

(19) Concomitant medications

(20) Adverse events evaluation

Follow-Up/Week 10, Visit 6 (Day 70±3 days).

The following procedures will be performed at this visit:

(1) Ongoing medical history review

(2) Vital signs

(3) Physical examination

(4) Safety labs (chemistry, coagulation, hematology and urinalysis):only if clinically significant change from baseline in safety labresults at week 8

(5) Urine pregnancy test (women of childbearing potential only)

(6) BSA evaluation

(7) IGA

(8) EASI assessment

(9) SCORAD assessment (including VAS pruritus assessment)

(10) POEM questionnaire

(11) DLQI questionnaire

(12) TEWL assessment (for selected sites only)

(13) Concomitant medications

(14) Adverse events evaluation

(15) Early termination visit.

In the case the subject ends the study before completion, the procedureslisted at Week 8 visit should be completed.

Study Assessments Efficacy Assessments

Clinical evaluations of atopic dermatitis will be performed by anexperienced and qualified dermatologist (board certified or equivalent).To assure consistency and reduce variability, the same assessor shouldperform all assessments on a given subject whenever possible.

Investigator's Global Assessment

The Investigator's Global Assessment (IGA) of Disease Severity (AppendixB) will be assessed at each visit. The IGA is a global assessment of thecurrent state of the disease. It is a 6-point morphological assessmentof overall disease severity and will be determined according to thefollowing definitions: 0 (clear), 1 (almost clear), 2 (mild), 3(moderate), 4 (severe) and 5 (very severe). In order to be eligible,subjects must have an IGA score>3 at Baseline visit (Day 0).

Eczema Area and Severity Index (EASI)

The Eczema Area and Severity Index (EASI) will be assessed at eachvisit, except screening visit. It quantifies the severity of a subject'satopic dermatitis based on both lesion severity and the percent of BSAaffected. The EASI is a composite score ranging from 0-72 that takesinto account the degree of erythema, induration/papulation, excoriation,and lichenification (each scored from 0 to 3 separately) for each offour body regions, with adjustment for the percent of BSA involved foreach body region and for the proportion of the body region to the wholebody. A detailed procedure of EASI score calculation is provided inAppendix C.

Body Surface Area (BSA)

The overall BSA affected by AD will be evaluated (from 0 to 100%) ateach visit. One patient's palm represents 1% of his/her total BSA. Forall study visits except at screening, the BSA of involved skin will bemeasured with the SCORAD measurement (see below for description) andevaluated as a separate endpoint. In order to be eligible, subjects musthave a BSA of at least 10% at Baseline visit (Day 0).

SCORing Atopic Dermatitis (SCORAD) SCORAD will be measured at eachvisit, except the screening visit. The SCORAD grading system wasdeveloped by the European Task Force on Atopic Dermatitis (1993) and hasbeen a standard tool to assess the AD severity in clinical studies inEurope. Six items (erythema, edema/papulation, oozing/crusts,excoriation, lichenification, and dryness) will be selected to evaluatethe AD severity. The overall BSA affected by AD will be evaluated (from0 to 100%) and included in the SCORAD scores. Loss of sleep and prurituswill be evaluated by patients on a visual analog scale (0-10). The sumof these measures represents the SCORAD which can vary from 0 to 103.The detailed procedure of SCORAD score calculation is provided inAppendix D.

Visual Analog Scale of Pruritus

For all study visits except screening, the pruritus severity score willbe recorded with the SCORAD measurement and this will be evaluated as aseparate endpoint. This will be evaluated by asking subjects to indicateon the 10-cm scale (0-10) of the assessment form the point correspondingto the average value for the last three days/nights.

Patient-Oriented Eczema Measure (POEM)

The Patient-Oriented Eczema Measure (POEM) will be assessed at eachvisit, except screening visit. The POEM developed by Charman et.al. is aself-assessment of disease severity by the patient. POEM has a maximumvalue of twenty eight based on the patient's response to seven questionsscored according to the following scale:

No Days=0

1-2 Days=1

3-4 Days=2

5-6 Days=3

Everyday=4

A detailed description of the POEM assessment is provided in Appendix E.

Dermatology Quality of Life (DLQI) Questionnaire

The DLQI is a simple 10-question validated questionnaire which will becompleted at each visit, except screening. The questionnaire is providedin Appendix F.

Transepidemal Water Loss (TEWL) (at Selected Sites Only)

The clinical severity of AD and associated effect on skin barrierfunction will be evaluated at each visit, except the screening visit.This evaluation will be performed at selected sites that havedemonstrated previous experience with this device.

At Baseline (Day 0), the investigator will select three representativeareas of active AD for each subject; the location of these sites will berecorded. At subsequent visits, TEWL readings for each area of AD willbe taken in standard room ambient conditions (22-25° C., 40-60% relativehumidity); the mean of the TEWL measurements will be used for theanalyses.

Safety Assessments Vital Signs

The following vital signs will be recorded at every visit in a seatedposition, after having sat calmly for at least 5 minutes: systolic anddiastolic blood pressure (mmHg), pulse (bpm), body temperature (° C.)and respiratory rate (breath/min).

Weight (kg) and Height (cm) will be collected to calculate the BMI, andwill be recorded at the Screening, Baseline and week 8 visits. Theheight will only be recorded once at the screening visit and the samevalue will be used for BMI calculation at baseline and Week 8 visits.

Any abnormal finding related to vital signs that the investigatorconsiders to be clinically significant, must be recorded as an AE.

Physical Examination

The following sites/systems will be included in the physicalexamination. Each system will be scored as normal/abnormal(non-clinically significant or clinically significant). Pertinentdetails must be recorded for any clinically significant findings.

(1) General appearance

(2) Dermatological (except Atopic dermatitis)

(3) Head, Eyes, Ears, Nose, Throat (HEENT)

(4) Respiratory

(5) Cardiovascular

(6) Abdominal

(7) Neurological

(8) Musculoskeletal

(9) Lymphatic

Clinical Laboratory Tests

Laboratory tests will be performed at screening, Day 0, Week 4 and Week8. If Week 8 results indicate a clinically significant change frombaseline, laboratory tests will also be performed at Week 10. The testswill include urinalysis, hematology with differential and coagulationtesting, a standard chemistry panel (chemistry includes liver functiontests and cholesterol), coagulation, serum pregnancy test (screening andweek 8/Early termination visits) for women of childbearing potential(WOCBP). At baseline (Day 0), week 2, week 4 and week 10 visits, a urinepregnancy test will be performed for women of childbearing potential(conducted at the investigator site). At screening visit, FSH levelswill be tested for women greater than 40 and less than 60 years of agewho have had a cessation of menses for at least 12 months but less than24 months. The specific tests in these panels are listed below in Table8:

TABLE 8 Clinical Laboratory Testing Laboratory testing Tests includedHematology Basophils, Eosinophils, HCT, HGB, Lympho- cytes, MCH, MCV,Monocytes, Neutrophils, platelets, RBC, WBC Coagulation APTT, INR, PTpanel (frozen) Serum Albumin, Alkaline Phosphatase, ALT, AST, ChemistryChloride, Cholesterol (non-fasting), CK, Creatinine (Enzymatic), GGT,Glucose Random, LDH, Potassium, Sodium, Total Bilirubin, Triglycerides,Urea (BUN), Uric Acid β-hCG for females of childbearing potential(Screening and week 8/Early termination) Urinalysis Blood, Glucose, pH,Protein Laboratory Tests FSH levels for women greater than 40 and lessthan Required at 60 years of age who have had a cessation of mensesScreening only for at least 12 months but less than 24 months.

Total and Free DGLA Plasma Levels

At Baseline, week 4 and week 8 visits, blood draws will be performedprior to study drug administration (no study drug administration at week8 visit). If a subject comes to the clinic after taking their daily doseof study medication, this subject will be required to come back thefollowing day for PK blood draws. Total DGLA and free DGLA trough plasmalevels will be measured. A second blood draw will be performed for laterevaluation of total fatty acid profile in plasma. The blood draw forserum chemistry analysis will be split in two aliquots for chemistryanalysis and later evaluation of interleukins.

The date and time of the subject's last dose at home before the visitwill be recorded accurately. The study site will instruct subjects notto take their daily study drug dose at home for week 4 visit. Dosingwill occur in the clinic during the study visit. The exact time of thesample collection must be recorded.

Blood samples will be processed as soon as possible, no later than 1hour after blood collection. The plasma obtained will be transferred inpolypropylene tubes. Each tube will be labeled in order to identify theanalyte to be assayed. All samples will be frozen in an uprightposition. The detailed instruction for PK, Total Fatty Acid andinterleukin sample collection, processing, storage and shipment will beprovided in the central laboratory manual. The labels on each tube willinclude at least the following information:

(1) Study protocol

(2) Site number

(3) Subject identification number

(4) Visit name

(5) Analyte name

(6) Primary (A) or Duplicate (B)

Shipment of the experimental samples will be shipped to ICON CentralLaboratories.

Detailed instructions for shipment will be provided in the centrallaboratory manual. Samples will then be shipped to the analyticalfacility and will be analyzed using a validated analytical method incompliance with their standard operating procedures.

Blood specimens for PK analysis will be maintained in a blinded fashion.

Adverse Events

An adverse event is any untoward medical occurrence in a patientadministered a pharmaceutical product, without regard to the possibilityof a causal relationship with this treatment.

Investigators are responsible for monitoring the safety of subjects whoare participating in this study and for alerting the Sponsor of anyevent that seems unusual, even if this event may be considered anunanticipated benefit to the subject. The investigator is responsiblefor appropriate medical care of subjects during the study.

The investigator remains responsible for following through anappropriate health care option, adverse events that are serious or thatcaused the subject to discontinue before completing the study. Thesubject should be followed until the event is resolved or stable.Follow-up frequency is left to the discretion of the investigator.

Safety will be evaluated by collecting adverse events, vital signs,performing physical examinations and evaluating laboratory results. Thereported adverse events will be coded according to MedDRA terminology.

Prior to enrollment, study site personnel will note the occurrence andnature of each subject's medical condition(s) in the appropriate sectionof the source document and CRF. During the study, site personnel willagain note any change in the condition(s) and the occurrence and natureof any adverse events.

If a subject experiences an adverse event after the first dose of thestudy drug, the event will be recorded as an adverse event in the sourcedocument and CRF. All AEs will be described in the source documents andin the CRF.

Adverse Events Causality

The investigator will establish causality of the AE to experimentaltreatment. The investigator should take into account the subject'shistory, most recent physical examination findings, and concomitantmedications.

The following definitions will be used to determine causality of an AE:

Not related: temporal relationship of the onset of the AE, relative tothe experimental treatment is not reasonable or another cause canexplain the occurrence of the AE.

-   Related: temporal relationship of the onset of the AE, relative to    the experimental treatment is reasonable, follows a known response    pattern to the treatment, and an alternative cause is unlikely.

Adverse Events Severity

The intensity of an AE is an estimate of the relative severity of theevent made by the investigator based on his or her clinical experienceand familiarity with the literature. The following definitions are to beused to rate the severity of an AE:

Mild: The symptom is barely noticeable to the subject and does notinfluence performance of daily activities. Treatment is not ordinarilyindicated.

Moderate: The symptom is sufficiently severe to make the subjectuncomfortable, and performance of daily activities is influenced.Treatment may be necessary.

Severe: The symptom causes severe discomfort, and daily activities aresignificantly impaired or prevented. Treatment may be necessary.

Serious Adverse Events

If a patient experiences a serious adverse event after the first dose ofthe study drug, the event will be recorded as a serious adverse event.All AEs will be described in the source documents and in the CRF.

A serious adverse event (experience) or reaction is any untoward medicaloccurrence that at any dose:

-   -   results in death,    -   is life-threatening,    -   requires in-subject hospitalization or prolongation of existing        hospitalization,    -   results in persistent or significant disability/incapacity, or    -   is a congenital anomaly/birth defect.

The term “life-threatening” in the definition of “serious” refers to anevent in which the subject was at risk of death at the time of theevent; it does not refer to an event which hypothetically might havecaused death if it were more severe.

Medical and scientific judgment should be exercised in deciding whetherexpedited reporting is appropriate in other situations, such asimportant medical events that may not be immediately life-threatening orresult in death or hospitalization but may jeopardize the subject or mayrequire intervention to prevent one of the other outcomes listed in thedefinition above. These should also usually be considered serious.

Examples of such events are intensive treatment in an emergency room orat home for allergic bronchospasm; blood dyscrasias or convulsions thatdo not result in hospitalization; or development of drug dependency ordrug abuse.

Pregnancy Reporting

If a subject becomes pregnant during the study, the subject shouldinform the study site as soon as possible. Upon confirmation of thepregnancy, the subject must be withdrawn from study drug but maycontinue study participation. The Investigator must complete astudy-specific Pregnancy Form upon confirmation of a pregnancy and sendit to Innovaderm Research within 24 hours of confirmation of thepregnancy. Innovaderm Research will report all cases of pregnancy to theSponsor in a timely manner (contact information to be used are the sameas for SAE reporting). Post-treatment follow-up should be done to ensuresubject safety. Pregnancy is not itself an AE or SAE; however,maternal/fetal complications or abnormalities will be recorded as AEs orSAEs, as appropriate. The investigator will follow the pregnancy untilcompletion (or until pregnancy termination) and notify InnovadermResearch of the outcome as a follow up to the initial Pregnancy Form.

Data Quality Assurance/Site Monitoring

During the study, monitoring visits will be conducted at regularintervals. The monitoring visits will be conducted to ensure protocoladherence, quality of data, accuracy of entries in the eCRF, drugaccountability, compliance with regulatory requirements and continuedadequacy of the investigational site and its facilities.

The site may be audited and/or monitored by a quality assurance officernamed by the Sponsor and/or regulatory authorities may wish to performon-site audits. The investigator will be given notice before an auditoccurs and will be expected to cooperate with any audit, provideassistance and documentation (including source data) as requested.

Data Collection and Retention

Subject data will be entered by site personnel using Medrio eClinicalOvernight, a web based electronic data capture (EDC) and reportingsystem. This application will be set up for remote entry. Medrio Inc.are the developers and owners of Medrio eClinical Overnight. The EDCsoftware has been fully validated and conforms to 21 CFR Part 11requirements. Investigator site staff will not be given access to theEDC system until they have been fully trained by the Sponsor ordelegate. Designated investigator staff will enter the data required bythe protocol into the eCRFs using this web based application. Automaticvalidation programs check for data discrepancies in the eCRFs and, bygenerating appropriate error messages, allow modification orverification of the entered data by the investigator staff beforeconfirming the data. The investigator must certify that the data arecomplete and accurate by applying an electronic signature to the eCRFs.

The investigator must maintain source documents for each subject in thestudy, consisting of case and visit notes (clinical medical records)containing demographic and medical information and the results for anytests or assessments. All information on the eCRFs must be traceable tothese source documents in the subject's file. Data not requiring awritten or electronic record will be defined before study start and willbe recorded directly on the eCRFs, which will be documented as being thesource data.

The data collected will be encoded and stored electronically in adatabase system. Validated data may subsequently be transferred to thesponsor.

Confidentiality of Trial Documents and Subject Records

The investigator must assure that the subjects' anonymity will bemaintained and that their identities are protected from unauthorizedparties. On CRFs or other document submitted to the Sponsor, subjectsshould not be identified by their names, but by an identification code.The Investigator should keep a subject enrolment log relating codes withthe names of subjects. The Investigator should maintain documents notfor submission to the sponsor e.g., subjects' written consent forms, inthe strictest confidence.

Investigator's Files/Retention of Documents

The Investigator must maintain adequate and accurate records to enablethe conduct of the study to be fully documented and the study data to besubsequently verified. These records include, but are not limited to,the identity of all participating subjects, all original signed informedconsent documents, copies of all CRFs, safety reporting forms, sourcedocuments, and detailed records of treatment disposition, and adequatedocumentation of relevant correspondence. These documents should beclassified into two different separate categories: Investigator StudyFile and Subject Clinical Source Documents.

The records should be retained by the investigator according toInternational Conference on Harmonisation (ICH), local regulations, oras specified in the Clinical Trial Agreement (CTA), whichever is longer.

Sample Size and Statistical Methods Determination of Sample Size

The primary endpoint can be translated as a responder analysis where asubject will be classified as Responder if he/she achieves an IGA scoreof 0 (clear) or 1 (almost clear) at Week 8, considering a 2-pointdecrease from baseline. A sample size of 45 subjects will have a powerof 80% to detect a statistically significant difference of 25% betweenresponders from treated group and from the placebo group, based on achi-square test and an alpha of 0.05. Based on the literature review, itis expected that the placebo could reach up to 7%, so the minimalproportion expected in the treated group should be at least 32%.Allowing for 10% drop-out, a total of 100 subjects should be enrolled inthe study.

Statistical and Analytical Plans

Continuous variables will be summarized in tables and will include thenumber of subjects, mean, standard deviation, median, minimum, maximumand inter-quartile range. Categorical variables will be presented intables as frequencies and percentages.

All statistical tests will be two-sided and will be performed with asignificant level of 0.05, unless otherwise specified.

Subject Disposition

Efficacy will be evaluated on the basis of the ITT population andanalyses will be performed based on the randomized treatment and not onthe treatment received.

The per-protocol (PP) population will include all subjects who wererandomized with no significant protocol deviations. The specificcriteria for the PP population and the ITT population will be detailedin a separate statistical analysis plan.

The safety population will be defined as all subjects who received atleast one dose of the medication. Analysis will be done according to theactual treatment they received.

Efficacy Analysis

The primary endpoint can be translated as a responder analysis where asubject will be classified as Responder if he/she achieves an IGA scoreof 0 (clear) or 1 (almost clear) at Week 8, considering a 2-pointdecrease from baseline. The comparison between groups for the primaryendpoint will be done using a chi-square test. A sensitivity analysis toexamine the contribution of site in the comparisons will be conductedusing a Cochran-Mantel-Haenszel test where the site will be used as thestratification factor. The primary efficacy analysis will be done usingobserved values and a supportive analysis will be conducted with atipping point analysis, a strategy used to evaluate the impact ofmissing values. Missing values will be set to responder/non-respondersuccessively and decisions based on p-values will be plotted. Details ofthis strategy will be presented in the statistical analysis plan. Therewill be no imputation for missing data at other visits. The analyseswill be done using the ITT population and will serve as the primaryanalysis while the analysis of the primary endpoint using the PPpopulation will be used a sensitivity analysis.

The secondary endpoints involving change from baseline will be analyzedusing an analysis-of-covariance (ANCOVA) including the change frombaseline as the dependent, the site and treatment group and site asfixed effects, and the baseline value as covariate. Ls-means and 95% Clwill be presented along with corresponding p-value from the comparisonof treatment. The secondary endpoints involving proportion will beanalyzed using a Cochran-Mantel-Haenszel test stratified by site andp-value will be presented. Analyses for the secondary endpoints will bedone using observed data and no imputation will be used for missingobservation.

All details regarding the statistical analyses will be included in aseparate statistical analysis plan.

Safety Analysis

All adverse events (AEs) that occur after the first study drugadministration during the study will be classified on the basis ofMedical Dictionary for Regulatory Activities (MedDRA) terminology.Descriptions of AEs will include the date of onset, the date the AEended (if it resolved), the severity and seriousness of the AE, therelationship of the AE to study medication, and the outcome. The focusin this protocol will be limited to treatment emergent adverse events.

Reported AEs will be summarized by the number of subjects reporting theevents, as well as by System Organ Class, Preferred Term, severity,seriousness, and relationship to study medication. For the summary ofAEs by severity, each patient will be counted only once within a SystemOrgan Class or a Preferred Term by using the AEs with the highestintensity within each category for each analysis. For the summary of AEsby relationship to study medication, each patient will be counted onlyonce within a System Organ Class or a Preferred Term by using the AEswith the greatest reported relationship within each category. For thesummary of AEs by relationship to study medication and severity, eachpatient will be counted only once within a System Organ Class or aPreferred Term by using (1) the greatest reported relationship followedby (2) the highest reported intensity.

All information pertaining to AEs noted during the study will be listedby patient, detailing verbatim, System Organ Class, Preferred Term,start date, stop date, intensity, outcome and relationship to studydrug. The AE onset will also be shown relative (in number of days) tothe day of test article administration. Serious adverse events (SAEs)will be tabulated by treatment group, relationship to the test article,and a reference to the occurrence of the SAEs to the relative day ofdosing.

Concomitant medications will be coded with the WHO-Drug Dictionary andlisted by subject.

In addition, a list of subjects who discontinued from the study and alist of subjects who experienced SAEs will also be provided.

Results from laboratory analyses and vital signs will be tabulated usingdescriptive statistics. The value at visit as well as the change frombaseline will be presented descriptively.

No inferential statistics will be done on safety variables (TEAEs,concomitant medication, laboratory and vital signs).

Ethics Local Regulations/Declaration of Helsinki

This study will be conducted in accordance with the ethical principlesthat have their origin in the Declaration of Helsinki (2008) and thatare consistent with “Good Clinical Practice” ICH Tripartite Guideline(July 2002) and the applicable laws and regulations of the country inwhich the research is conducted, whichever affords the greaterprotection to the individual.

Ethical Review

It is the understanding of the Sponsor that this protocol (and anyamendment) as well as appropriate consent procedures, will be reviewedand approved by a research ethics board/institutional review board(REB/IRB). This Board must operate in accordance with the currentFederal regulations. A letter or certification of approval will be sentby the Investigator to the Sponsor prior to initiation of the study, andalso whenever subsequent modifications to the protocol are made.

Informed Consent

It is the responsibility of the Investigator, or a person designated bythe Investigator (if acceptable by local regulation), to obtain writteninformed consent from each individual participating in this study afteradequate explanation of the aims, methods, objectives and potentialhazards of the study. It must also be explained to the subjects thatthey are completely free to refuse to enter the study or to withdrawfrom it at any time for any reason.

If new safety information results in significant changes in therisk/benefit assessment or any new information that may affectwillingness to continue to participate, the consent form should ifnecessary be reviewed and updated by the Research EthicsBoard/Institutional Review Board. All subjects (including those alreadybeing treated) should be informed of the new information, given a copyof the revised form and asked to give their consent to continue in thestudy.

APPENDIX A Study Flow Chart

Visit Week 8/ Follow up Screening Baseline Week 2 Week 4 ET visitvisit/Week 10 Day −30 to −1 0 14 (±2) 28 (±2) 56 (±2) 70 (±3) InformedConsent X Demographics X Medical/Surgical History X X Ongoing medicalhistory

X X Review Inclusion/Exclusion Criteria X X Hanifin and Rajka criteria XAssign subject identifier number X Randomization X ConcomitantMedications X X X X X X Safety labs and inflammation X^(Φ) X X X(X**)^(Φ) biomarkers: Serum Chemistry (including FSH levels at screeningwhen applicable^(¥) and interleukin profile), Coagulation, Hematology,urinalysis Pharmacokinetics (pre-dose blood X X X draw)^(††) Blood drawfor fatty acid profile X X X sample^(††) Pregnancy Test (β-hCG if femaleof X X X X X X childbearing potential)* Vital Signs X X X X X X PhysicalExamination X X X X BMI X X X Study Drug Administration (on site)

X X ^(§) Dispense Study Drug X X X Collect Study Drug X X X DispenseSubject Compliance Log X X X Collect and Review Subject X X X ComplianceLog Capsule count X X X BSA X X X X X X IGA X X X X X X EASI assessmentX X X X X SCORAD assessment/VAS X X X X X pruritus assessment POEMquestionnaire X X X X X DLQI questionnaire X X X X X TEWL (selectedsites only) X X X X X Adverse Events^(†) X^(†) X X X X ^(¥)For womengreater than 40 and less than 60 years of age who have had a cessationof menses for at least 12 months but less than 24 months **Only ifclinically significant change from baseline in safety lab results atweek 8 *Serum pregnancy test at screening and week 8/ET visits, urinetest pregnancy for all other visits ^(††)If a subject took studymedication prior to the visit, he/she will be required to come back thefollowing day for PK blood draws.

Subjects must be fasting for at least 8 hours before and 60 minutesafter drug administration ^(§)Subjects will be instructed to take theirlast study drug dose the day preceding week 8 visit. ^(†)Collection ofAE will start after the first study drug administration

Assessment if any ongoing condition has improved since baseline.^(Φ)Interleukin profile will not be evaluated at screening and week 10.

APPENDIX B—Investigator's Global Assessment

Score Grade Definition 0 Clear No evidence of disease with the exceptionof residual pigment changes and/or xerosis 1 Almost clear Perceptibleerythema, papulation/infiltration 2 Mild Mild erythema,papulation/infiltration 3 Moderate Moderate erythema,papulation/infiltration 4 Severe Severe erythema,papulation/infiltration 5 Very Severe Severe erythema,papulation/infiltration with oozing/crusting

APPENDIX C—Eczema Area and Severity Index (EASI)

Four anatomic sites—head, upper extremities, trunk and lowerextremities—are assessed for erythema, induration (papules), excoriationand lichenification as seen on the day of the examination. The severityof each sign is assessed using a 4-point scale:

0=No symptoms

1=Slight

2=Moderate

3=Marked

The area affected by atopic dermatitis within a given anatomic site isestimated as a percentage of the total area of that anatomic site andassigned a numerical value according to the degree of atopic dermatitisinvolvement as follows:

0=no involvement

1=<10%

2=10 to <30%

3=30 to <50%

4=50 to <70%

5=70 to <90%

6=90 to 100%

The EASI score is obtained by using the formula

EASI=0.1(Eh+Ih+Exh+Lh)Ah+0.2(Eu+Iu+Exu+Exu)Au+0.3(Et+It+Ext+Ext)At+0.4(El+Il+Exl+Exl)Al

Where E, I, Ex, L and A denote erythema, induration, excoriation,lichenification and area, respectively, and h, u, t, and l denote head,upper extremities, trunk, and lower extremities, respectively.

APPENDIX D—SCORing Atopic Dermatitis (SCORAD)

Six items (erythema, edema/papulation, oozing/crusts, excoriation,lichenification, and dryness) are selected to evaluate the AD severity.The intensity of each item is graded using a 4-point scale:

0=No symptoms

1=Mild

2=Moderate

3=Severe

The area chosen for grading must be representative (average intensity)for each item. The individual intensity ratings for each item will thenbe added (ranging from 0-18) and multiplied by 3.5, giving a maximalscore of 63.

The overall BSA affected by AD is evaluated (from 0 to 100%) and dividedby 5. One patient's palm represents 1% of his/her total BSA. The maximumis 20.

Subjective items include loss of sleep and the occurrence of pruritus.These are evaluated by asking patients to indicate on the 10-cm scale(0-10) of the assessment form the point corresponding to the averagevalue for the last three days/nights. The combined maximum score ofthese two is 20.

The sum of the above measures represents the SCORAD which can vary from0 to 103. If the subjective scores of pruritus and loss of sleep areexcluded, the SCORAD becomes objective SCORAD (score range 0-83).

APPENDIX E—Patient-Oriented Eczema Measure (POEM)

Patient ID #:      -          Patient Initials:          Visit Day:         Visit Date (dd-mmm-yyyy):           Please circle one responsefor each of the seven questions below about your eczema. Please leaveblank any questions you feel unable to answer. Please circle oneresponse for each of the seven questions below about your eczema. Pleaseleave blank any questions you feel unable to answer. 1. Over the lastweek, on how many days has you skin been itchy because of your eczema?No days 1-2 days 3-4 days 5-6 days Every day 2. Over the last week, onhow many nights has your sleep been disturbed because of your eczema? Nodays 1-2 days 3-4 days 5-6 days Every day 3. Over the last week, on howmany days has your skin been bleeding because of your eczema? No days1-2 days 3-4 days 5-6 days Every day 4. Over the last week, on how manydays has your skin been weeping or oozing clear fluid because of youreczema? No days 1-2 days 3-4 days 5-6 days Every day 5. Over the lastweek, on how many days has your skin been cracked because of youreczema? No days 1-2 days 3-4 days 5-6 days Every day 6. Over the lastweek, on how many days has your skin been flaking off because of youreczema? No days 1-2 days 3-4 days 5-6 days Every day 7. Over the lastweek, on how many days has your skin felt dry or rough because of youreczema? No days 1-2 days 3-4 days 5-6 days Every day

©C R Charman, A J Venn, H C Williams, December 2004.

APPENDIX F—Dermatology Life Quality Index (DLQI)

The aim of this questionnaire is to measure how much your skin problemhas affected your life OVER THE LAST WEEK. Please check one box for eachquestion.

1. Over the last week, how itchy, sore, painful or stinging has yourVery much □ skin been? A lot □ A little □ Not at all □ 2. Over the lastweek, how embarrassed or self conscious have Very much □ you beenbecause of your skin? A lot □ A little □ Not at all □ 3. Over the lastweek, how much has your skin interefered with you Very much □ Notrelevant □ going shopping or looking after your home or yard? A lot □ Alittle □ Not at all □ 4. Over the last week, how much has your skininfluenced the Very much □ Not relevant □ clothes you wear? A lot □ Alittle □ Not at all □ 5. Over the last week, how much has your skinaffected any social Very much □ Not relevant □ or leisure activities? Alot □ A little □ Not at all □ 6. Over the last week, how much has yourskin made it difficult for Very much □ Not relevant □ you to do anysport? A lot □ A little □ Not at all □ 7. Over the last week, has yourskin prevented you from working or yes □ Not relevant □ studying? no □If “No”, over the last week how much has your skin been a A lot □problem at work or studying A little □ Not at all □ 8. Over the lastweek, how much has your skin created problems Very much □ Not relevant □with your partner or any of your close friends or relatives? A lot □ Alittle □ Not at all □ 9. Over the last week, how much has your skincaused any sexual Very much □ Not relevant □ difficulties? A lot □ Alittle □ Not at all □ 10. Over the last week, how much of a problem hasthe treatment for Very much □ Not relevant □ your skin been, for exampleby making your home messy, or by A lot □ taking up time? A little □ Notat all □

The aim of this questionnaire is to measure how much your skin problemhas affected your life OVER THE LAST WEEK. Please check one box for eachquestion.

APPENDIX G—Diagnostic Criteria for Atopic Dermatitis

Per Inclusion Criterion 2, a subject is to have a clinical diagnosis ofatopic dermatitis according to the criteria of Hanifin and Rajka. Thecriteria go as follows:

Major Criteria (Must Have at Least Three)

Pruritus

Typical morphology and distribution:

Adults: flexural lichenification or linearity

Children and infants: involvement of facial and extensor surfaces

Chronic or relapsing dermatitis

Personal or family history of atopy

Minor Criteria (Must Have at Least Three)

Xerosis

Icthyosis/keratosis pilaris/palmer hyperlinearity

Immediate (type 1) skin test reactivity

Elevated serum IgE

Early age at onset

Tendency to skin infections (Staphylococcus aureus, herpessimplex)/impaired cellular

Immunity

Hand/foot dermatitis

Nipple eczema

Conjunctivitis

Dennie-Morgan fold

Keratoconus

Anterior subcapsular cataracts

Orbital darkening

Facial pallor/erythema

Pityriasis alba

Anterior neck folds

Itch when sweating

Intolerance to wool and lipid solvents

Perifollicular accentuation

Food intolerance

Course influenced by environmental/emotional factors

White demographic/delayed blanch

Example 3

An in vivo investigation of anti-hypertension efficacy followingco-administration of DGLA and aspirin in spontaneous hypertensive ratswas conducted. This experiment investigated the efficacy of DGLA whenco-administered chronically with aspirin to reduce the hypertensiveresponse induced acutely by phenylephrine in spontaneous hypertensiverats. Spontaneously hypertensive adult male rats (250-300 g), bred byCharles River Laboratories, were used for this assay. The animals wereidentified upon arrival as per CCPA guidelines. The animals werepair-housed by group (low dose+Aspirin, high dose+Aspirin or Aspirinalone) for the duration of the oral gavage prior to surgery. All animalcare and vivarium maintenance was recorded, with documents kept at thetest facility.

The rat is a well-characterized and highly sensitive model forassessment of vascular tension effects and evaluation of efficacyfollowing chronic exposure to a test article. This study design is basedon current International Conference on Harmonization (ICH) HarmonizedTripartite Guidelines [ICH S7A] and generally accepted procedures forthe testing of pharmaceutical compounds. This nonclinical laboratorystudy was designed as a non-GLP compliant study and did not require QAinvolvement.

The concentrations of DGLA to be tested (50 and 500 mg/kg) as well asthe concentration of aspirin (10 mg/kg) were selected by the Sponsorbased on information available at the time of design of this study. Theywere selected to cover a range of concentrations useful for theidentification of the mechanism of action of the test article.

The concentrations of positive control (carvedilol 0.1 mg/kg) and thehypertensor (phenylephrine 3, 10, and 20 μg/kg) were selected based onliterature references. All solution preparations were documented on aSolution Contents Form, defining labelling information and all relevantinformation on reagents: batch number, storage conditions, contents, andpreparation/expiry dates.

The test article was formulated using olive oil as the vehicle. Thisstock solution was then administered by oral gavage once per day forseven consecutive days. A low and a high dose (50 mg/kg and 500 mg/kg)were tested.

A 5 mg/ml stock solution of Aspirin was prepared by dissolving theappropriate amount of aspirin in 100% DMSO which was then diluted inwater. The concentration of DMSO was less than 1%. The appropriatevolume was administered by oral gavage to each animal once per day forseven consecutive days. The stock solution was stored at roomtemperature and was considered stable for the duration of the gavage.

A 0.4 mg/ml stock solution of carvedilol was prepared in PBS(pH=4.00±0.05). The appropriate volume was administered to each animalby an intravenous injection at the end of the experiment on the surgeryday. The stock solution was stored at 4° C. and was considered stablefor the duration of the experiment.

A stock solution of phenylephrine of 1 mg/ml were prepared in PBS(pH=7.4±0.05). The stock solution was stored at 4° C. The expirationdate was set at 14 days after preparation.

Any remaining test article will be destroyed after completion of thestudy.

Male spontaneous hypertensive rats weighing approximately 300 g arrivedat the facility at least two days before beginning the oral gavage.Animals were assigned to one of three groups (low dose+aspirin, highdose+aspirin, and aspirin), and were pair-housed during the acclimationperiod. Each group (n=4) received the appropriate compound by oralgavage using a 16G feeding needle. The stock solution was administeredonce per day for seven consecutive days. On the seventh day ofadministration, the animals underwent surgery and hypertensivemonitoring.

Rats were anesthetized with a 2.5-3.0% isoflurane-O2 mixture in aninduction box. When the animals were removed from the induction box,they were connected onto a nose cone to maintain anesthesia during thetracheotomy. The animals were tracheotomised with an endotracheal tube(7 cm length from connector to the tip made with a PE205 tube from BDand a 16G needle) to facilitate spontaneous breathing, stabilizehemodynamics, and to keep the rat under anesthesia with an isoflurane-O2mixture. Two catheters were inserted: one in the right femoral arteryfor systemic arterial pressure (SAP) measurement, and one in the rightfemoral vein for delivery of the hypertensor and positive control. ECGsignals were acquired with ECG contacts placed in a Lead-1 configurationon the anaesthetized animal, and a pulse oxymeter was attached to theanimal's hind paw to enable continuous monitoring of the generalcondition of the rat during the surgical. Following baselinemeasurements, three doses of phenylephrine (3, 10 and 20 μg/kg) weredelivered by an IV bolus injection, leaving 5 minutes between doses. Onedose of the positive control, carvedilol (0.1 mg/kg), was administered,following the highest concentration of phenylephrine. After the positivecontrol, 10 μg/kg of phenylephrine was administered again. Bloodpressure, systemic arterial pressure (SAP), and heart rate weremonitored continuously for a total of half an hour. Hypertension wasdetected as an increase in diastolic, systolic or mean blood pressure.At the end of the experiment, the animals were euthanized byexsanguination.

The analysis software used was Clampfit 10.2.0.14 by Axon Instruments,installed on networked personal computers running Microsoft Windows.Clampfit 10.2.0.14 has been fully validated in the connected context inwhich it is used. The graphics software for illustrations is MicrosoftOffice Excel 2007 installed on networked personal computers runningMicrosoft Windows. The continuously recorded systemic arterial pressure(SAP) was used to calculate the mean systemic arterial pressure (mSAP)for each condition. The heart rate was monitored continuously from thetime anesthesia was induced. One-way ANOVAs, comparing pre- andpost-exposure parameters across experimental groups, were analysed.Statistical significance was confirmed at p≦0.05.

Table 9. Arterial pressure and mean systemic arterial pressure (mm Hg)following seven days of gavage, before intravenous doses ofphenylephrine.

TABLE 9 Mean systemic Treatment Systol/Diastol arterial pressure OliveOil (vehicle) 160/100 120 DGLA 50 mg/kg 120/75  90 DGLA 500 mg/kg135/80  98 Aspirin 10 mg/kg 190/100 130 DGLA 50 mg/kg + Aspirin 10 mg/kg180/120 140 DGLA 50 mg/kg + Aspirin 10 mg/kg 180/120 140 *Compared tobaseline ** Compared to PHE 20 μg/kg † Compared to second baseline §Compared to Carvedilol

Table 10 shows change in mean systemic arterial pressure withintravenous phenylephrine doses following seven consecutive days ofgavage with Aspirin at 10 mg/kg/day.

TABLE 10 Treatment   Aspirin 10 mg/kg Mean Arterial Pressure AverageTest vs Conditions Rat #1 Rat #2 Rat #3 Rat #4 Rat #5 (mmHg) STED SEMbaseline n Baseline 86.75 107.64 160.50 141.36 187.84 136.8 40.44 18.09N/A 5 PHE 3 μg/kg 97.24 122.18 168.06 150.54 192.06 146.0 * 37.33 16.690.006 5 PHE 10 μg/kg 137.63 122.18 203.75 169.61 201.69 167.0 * 36.8516.48 0.016 5 PHE 20 μg/kg 149.35 120.84 202.61 194.76 203.90 174.3 *37.34 16.70 0.019 5 Baseline 98.38 64.32 164.88 159.32 177.61 132.9 **49.03 21.93 0.002 5 Carvedilol 0.1 mg/kg 57.12 44.38 145.56 134.82115.06 99.4_(†) 45.95 20.55 0.015 5 PHE 10 μg/kg 93.22 52.61 185.63171.45 187.96 138.2_(§) 61.60 27.55 0.020 5

FIG. 1 illustrates the change in mean systemic arterial pressure, in mmof Hg, following phenylephrine administration. Five rats receivedintravenous doses of phenylephrine after seven consecutive days ofgavage with Aspirin at 10 mg/kg/day. The mean systemic arterial pressureat each of the three doses of phenylephrine was compared statisticallyto the first baseline, while the carvedilol condition was statisticallycompared to the baseline immediately prior to administration, and thefinal dose of phenylephrine (10 μg/kg) was compared to the positivecontrol condition directly before.

These results demonstrate an increased mean systemic arterial pressurewith increasing doses of phenylephrine. All data obtained werestatistically different from baseline, confirming the hypertensor effectof phenylephrine in the spontaneously hypertensive rat model fed withaspirin. The effect of phenylephrine was fully reversible; the meanarterial pressure during the second baseline (following 20 μg/kg) wasactually quite similar to the original baseline.

Carvedilol caused a significant decrease when added immediately afterphenylephrine, confirming its well-known ability to reduce high bloodpressure (carvedilol is a non-specific beta blocker/alpha-1 blocker andblocks the binding of phenylephrine to alpha-1 adrenergic receptors).The last addition of phenylephrine, following the carvedilol, caused alimited increase in the mean systemic arterial pressure.

FIG. 2. Change in mean arterial pressure (mmHg) with intravenous dosesof phenylephrine following seven consecutive days of gavage with DGLA at50 mg/kg+Aspirin at 10 mg/kg.

Table 11. Change from baseline in mean arterial pressure withintravenous phenylephrine doses following seven consecutive days ofgavage with DGLA at 50 mg/kg+Aspirin at 10 mg/kg.

TABLE 11 Treatment   50 mg/kg + Aspirin 10 mg/kg Mean Arterial PressureTtest Average vs Conditions Rat #1 Rat #2 Rat #3 Rat #4 Rat #5 SAP STEDSEM Aspirin n Baseline 136.52 148.19 176.99 108.14 77.13 129.4 38.2517.11 0.773 5 PHE 3 μg/kg 154.55 154.66 185.15 117.40 81.18 138.6 40.0817.93 0.770 5 PHE 10 μg/kg 192.63 188.72 193.26 130.48 94.43 159.9 45.1920.21 0.793 5 PHE 20 μg/kg 196.59 196.00 196.67 148.23 141.78 175.928.25 12.64 0.942 5 Baseline 169.28 160.97 160.79 94.77 78.64 132.942.68 19.09 1.000 5 Carvedilol 0.1 mg/kg 115.29 97.01 126.33 59.10 58.9391.3_(†) 31.30 14.00 0.006 5 PHE 10 μg/kg 178.71 160.97 n/d 86.29 74.07125.0_(§) 52.51 26.25 0.043 4

The results presented in FIG. 2 demonstrate changes in mean arterialpressure, in mm of Hg, after intravenous doses of phenylephrine in fivespontaneously hypertensive rats following seven days of oral feedingwith DGLA at 50 mg/kg+Aspirin at 10 mg/kg. The results for the testarticle were compared to the data obtained with the aspirin only group(10 mg/kg). Carvedilol was compared to the second baseline, and the lastdose of phenylephrine was statistically compared to the carvedilol.

The animals exhibited a dose-dependent increase in blood pressure asphenylephrine concentration was increased. The rats which had beenadministered DGLA 50 mg/kg and aspirin exhibited an increase in arterialpressure for a given dose of phenylephrine which was essentially thesame as that measured in the rats having received Aspirin over 10 days.With the rats which were fed with DGLA at 50 mg/kg in addition of 10mg/kg of aspirin, there was no significant difference in the meanarterial pressure when compared to the aspirin group at any of theconcentrations of phenylephrine. It would thus appear that Aspirin dailyprevents the benefits obtained from daily doses of 50 mg/kg DGLA.

Carvedilol caused a significant decrease in blood pressure in DGLA+aspirin-fed animals. Due to a problem with the anesthesia, rat #3 wasnot given the last dose of phenylephrine (n=4 for this condition).

Table 12. Change in mean arterial pressure with intravenousphenylephrine doses following seven consecutive days of gavage with DGLAat 500 mg/kg co-administered with Aspirin at 10 mg/kg.

TABLE 12 Treatment   500 mg/kg + Aspirin 10 mg/kg Mean Arterial PressureAverage Ttest vs Conditions Rat #1 Rat #2 Rat #3 Rat #4 SAP STED SEMAspirin n Baseline 145.62 100.15 128.35 129.60 125.9 18.90 9.45 0.638 4PHE 3 μg/kg 161.09 103.72 136.18 133.59 133.6 23.49 11.74 0.584 4 PHE 10μg/kg 170.69 111.08 145.53 149.12 144.1 24.66 12.33 0.325 4 PHE 20 μg/kg161.17 116.35 141.85 140.72 140.0 18.36 9.18 0.140 4 Baseline 120.1683.43 88.30 85.82 94.4 17.27 8.64 0.182 4 Carvedilol 0.1 mg/kg 62.1357.56 57.32 52.11 57.3_(†) 4.10 2.05 0.014 4 PHE 10 μg/kg 95.50 65.1980.39 58.97 75.0 16.36 8.18 0.070 4

FIG. 3 illustrates the changes in mean arterial pressure followingintravenous doses of phenylephrine after seven consecutive days ofgavage with DGLA 500 mg/kg co-administered with aspirin at 10 mg/kg infour spontaneously hypertensive rats. As with the lower dose ofDGLA+aspirin, the phenylephrine conditions were compared to those of theaspirin only group. Carvedilol was statistically compared to thebaseline just before administration, while the last dose ofphenylephrine (10 μg/kg) was compared to the positive control.

Despite lower mean arterial pressures observed, there was no significantstatistical difference between the mean systemic arterial pressure ofthe rats which were fed with both DGLA and aspirin and those who werefed with aspirin only. It would thus appear that Aspirin again preventedthe benefit of daily administration of 500 mg/kg DGLA. The positivecontrol succeeded in statistically decreasing the arterial bloodpressure of the animals, as observed in the other groups. The increasecaused by the last dose of phenylephrine was not considered significant.

Table 13 shows mean arterial pressure at baseline following sevenconsecutive days with six different gavage groups.

TABLE 13 Mean Arterial pressure at baseline Average Conditions Rat #1Rat #2 Rat #3 Rat #4 Rat #5 SAP STED SEM Ttest n Vehicle 70.64 77.31120.52 93.60 n/a 90.5 22.21 11.10 n/a 4 DGLA 50 mg/kg 51.77 87.81 62.1186.01 84.93 74.5 16.50 7.38 n/a 5 DGLA 50 mg/kg + 136.52 148.19 176.99108.14 77.13 129.4* 38.25 17.11 0.019 5 Aspirin DGLA 500 mg/kg 64.8191.55 94.41 66.56 93.64 82.2 15.12 6.76 n/a 5 DGLA 500 mg/kg + 145.62100.15 128.35 129.60 n/a 125.9* 18.90 9.45 0.006 4 Aspirin Aspirin 86.75107.64 160.50 141.36 187.84 136.8 40.44 18.09 n/a 5

FIG. 4 presents the mean arterial pressures at baseline of the sixgroups which were fed during the two parts of this study. Thetracheotomy stabilized hemodynamics, but also caused changes in thearterial pressure of the rats. The arterial pressure of the rats who didnot receive aspirin (Vehicle, DGLA 50 mg/kg and DGLA 500 mg/kg) wereobtained during the first phase of this study (study #20131022-1) andthose of the other groups were recorded during this present study(different batches of animals from Charles River, but same strain andsize of rat). Statistical t tests were done between the groups whichwere administered DGLA at 50 or 500 mg/kg and their equivalent addedwith aspirin at 10 mg/kg. There was a statistically significant increaseof the mean arterial pressure at baseline between the 50 mg/kg of DGLAalone and the 50 mg/kg of DGLA co-administered with aspirin at 10 mg/kg,as well as between the 500 mg/kg group of DGLA compared to DGLA 500mg/kg with aspirin 10 mg/kg (significant increase marked with *).

It would thus appear that DGLA successfully decreases the arterialpressure in spontaneously hypertensive rats, an effect which iseliminated by Aspirin.

Table 14 shows mean arterial pressure with an intravenous dose ofphenylephrine at 20 μk/kg following seven consecutive days with sixdifferent gavage groups.

TABLE 14 Mean Arterial pressure with an intravenous dose ofPhenylephrine 20 μg/kg Average Conditions Rat #1 Rat #2 Rat #3 Rat #4Rat #5 SAP STED SEM Ttest n Vehicle 131.99 130.56 145.47 137.32 n/a136.3 6.75 3.37 n/a 4 DGLA 50 mg/kg 91.82 137.84 105.44 132.68 110.72115.7 19.23 8.60 n/a 5 DGLA 50 mg/kg + 196.59 196.00 196.67 148.23141.78 175.9* 28.25 12.64 0.004 5 Aspirin DGLA 500 mg/kg 105.21 119.33118.12 97.17 120.82 112.1 10.43 4.66 n/a 5 DGLA 500 mg/kg + 161.17116.35 141.85 140.72 n/a 140.0* 18.36 9.18 0.023 4 Aspirin Aspirin149.35 120.84 202.61 194.76 203.90 174.3 37.34 16.70 n/a 5

FIG. 5 presents the mean arterial pressures of all the groups analyzedover the two studies investigating DGLA. These arterial pressures wereobtained after an intravenous dose of phenylephrine at 20 μg/kg. As withthe results acquired in baseline, each dose of DGLA was statisticallycompared to its equivalent condition in the aspirin only group. Asignificant augmentation of the mean arterial pressure was noticedbetween both corresponding groups (marked with *).

This study was designed to investigate the efficacy of DGLA, alone andwhen co-administered chronically with aspirin, to reduce the arterialpressure measured in hypertensive rats, as well as the reduction in thehypertensive response induced acutely by phenylephrine in the samestrain of spontaneously hypertensive rats.

The Spontaneously Hypertensive rat (SHR) is a commonly used model inhypertension research because it produces, within each colony, uniformchanges in response to direct and indirect effectors to thecardiovascular system. The rats have been selected and inbred overgenerations, defined as hypertensive by a systolic blood pressure ofover 150 mm Hg persisting for more than one month.

The study revealed that the chronic co-administration of DGLA andaspirin prevented DGLA from lowering the mean systemic arterial bloodpressure (by up to 25%) as observed previously in the spontaneoushypertensive rats treated with the DGLA.

The doses of DGLA tested (50 and 500 mg/kg) combined with aspirin at 10mg/kg did not significantly reduce the dose-dependent elevation of themean arterial blood pressure following phenylephrine injection, whencompared to animals which were fed with aspirin alone. The two doses ofDGLA caused similar effects, suggesting that the benefits of greaterdoses of DGLA may be minimal if Aspirin is co-administered. When bothdoses of DGLA were compared to their equivalent aspirin-only conditions,Aspirin caused a clear increase in the mean arterial pressure inbaseline condition as well as when the rats were challenged with anintravenous dose of 20 μg/kg of phenylephrine.

In this study, Carvedilol was used as the positive control. It is abeta-blocker used to treat heart failure and high blood pressure. Itacts by relaxing blood vessels and slowing heart rate, thus improvingventricular refilling, blood flow, and decreasing blood pressure. Theeffect of carvedilol, when injected intravenously in spontaneouslyhypertensive rats, confirmed its ability to decrease blood pressureafter each of three doses of phenylephrine, and demonstrated thesensitivity of the test system.

Example 4

A single-center, double-blind, randomized, placebo-controlled, two partPhase one study was performed in healthy male and female subjects aged18 to 45 years inclusive. The primary objective of this study was toassess the safety and plasma pharmacokinetics (PK) of orallyadministered single doses and orally administered multiple doses ofDS107G capsules (once daily for 28 days) in healthy subjects.

The secondary objectives of this study were to assess the following:

The effect of food on the PK of an orally administered single dose ofDS107G capsule in healthy subjects.

The PK of DGLA in human skin following multiple doses of DS107G capsulegiven once daily for 28 days to healthy subjects.

A total of 48 subjects were enrolled (including 32 for single dose studyand 16 for multiple dose study). The single and multiple dose studiesconsist of 8 subjects per cohort (ideally 4 males and 4 females, but nofewer than 3 per sex).

Subjects were randomly assigned in a 3:1 ratio of DG107G to placebo byblock randomization. Single dose part one of the study comprised threecohorts of eight subjects each, with the addition of Cohort four basedon evaluation of the safety data of Cohort three by the SafetyMonitoring Committee (SMC). A single oral dose of DS107G or matchingplacebo was administered to fasted subjects in Cohorts one to three(500, 1000, and 2000 mg, respectively) in parallel; subjects in Cohortfour were administered a dose of 4000 mg following completion of Cohortthree. Subjects in Cohort two received 1000 mg or matching placebo inthe fed state at ≧14 days after the first dose.

Multiple dose part two of the study comprised two cohorts (Cohorts fiveand six) of eight subjects each, who were administered multiple doses ofstudy drug in the fasted state for 28 days. Subjects in Cohorts five andsix received DS107G 2000 or 4000 mg, respectively, or matching placeboonce daily for 28 days. Cohort six was started after Cohort five safetydata were evaluated by the SMC. No interim analysis was planned.

In Part one of the study, plasma PK analysis of Dihomo-Gamma-LinolenicAcid (DGLA) and 15-Hydroxyeicosatrienoic acid (15-HETrE), in fastedsubjects alone, was performed for Cohorts one to four up to 312 hourspost dose on study Days 1 to 5, 8, 14, and the follow-up visit. In Parttwo, plasma PK analysis of DGLA was performed for Cohorts five and sixup to 168 hours post dose on study Days 1, 2, 3, 5, and 8. Safetyassessments were monitored throughout the study.

A randomized, placebo-controlled, double blind design was used tominimize bias during the safety and tolerability assessments.

Part one of the study followed a single ascending dose design with adose escalation to 4000 mg. With the exception of the 4000-mg dose, allof the doses in Part one of the study were previously tested in humans.Treatment-related adverse events (TEAEs) were not observed in thepreviously tested multiple-dose regimens such as 150 mg/day for 28 days,450 mg/day for 28 days, 1000 mg/day for 14 days, and 2000 mg/day for 10days. The dose selection was based on the doses of oral DGLA tested inprevious clinical studies and the characterization of the PK and safetyof previously tested and higher doses were observed as main objective.

The effect of food on the oral bioavailability of DGLA was alsoassessed.

Healthy female subjects were included in this study to assess apotential gender component on the biomarker di-hydrotestosterone as DGLAis involved in steroid metabolism.

The total duration of participation for each subject in Part one of thestudy was approximately 14 days, excluding the screening period. In Part2 of the study, the duration of was approximately 42 days.

Screening procedures were conducted for both parts one and two beforethe start of the study on Day −1 (Baseline) by evaluating the safetyassessments. Safety assessments included adverse events (AEs), clinicallaboratory testing (hematology, biochemistry, virology [hepatitis Bsurface antigen, human immunodeficiency virus (HIV) antibodies,hepatitis C antibodies], and urine analysis), drug of abuse (DOA) testresults, and pregnancy test for female subjects (urine β human chorionicgonadotropin [βHCG]), vital signs (blood pressure [BP], pulse,temperature), 12-lead electrocardiograms (ECGs), physical examinations,and assessment of concomitant medications (only part two of the study).

All subjects were required to have met the inclusion and exclusioncriteria described below. However, minor deviations that were clinicallyinsignificant and that posed no safety concerns had been deemedacceptable by the Investigator and the Sponsor, consistent with theprotocol.

All subjects considered for study participation were required to havemet the following inclusion criteria:

-   1. Subject was male or female.-   2. Female subject and female partner of male subject:    -   Must not have been pregnant (female subjects must have had a        negative urine pregnancy test prior to entry into the study).    -   Must not have been breast feeding.    -   Must not have been planning to become pregnant during the study        period or within 3 months following the study.    -   Must have been adhering to an adequate form of contraception        prior to entry into the study and for an additional 3 months        after the follow-up visit.-   3. Subject was aged between 18 and 45 years inclusive.-   4. Subject had signed the informed consent form.-   5. Subject's body mass index (BMI) was between 18.0 and 30.0 kg/m²    inclusive.-   6. Subject was considered to be in good health in the opinion of the    Investigator by evaluating the safety assessments.-   7. Subjects must have been able to communicate well with the    Investigator, to understand and comply with the requirements of the    study, and to understand and sign the written ICF. Subjects were    excluded from the study if there was evidence of any of the    following criteria: Subject had a clinically significant illness in    the 4 weeks before screening.-   8. Females of childbearing potential and female partners of male    subjects who had not used a safe contraceptive measure for 3 months    prior to the study and were not willing to use a safe contraceptive    measure for the duration of the study; examples of a safe    contraceptive measure included intra-urine device or oral    contraceptives, diaphragm, or condom if used in combination with a    spermicide.-   9. Subject used a prescribed medication in the 2 weeks before dosing    or used over-the-counter preparations (including vitamins and    supplements) for 1 week before dosing (with the exception of    paracetamol, which was allowed up to 48 hours before dosing), and    hormonal contraceptives.-   10. Subject used dietary supplements rich in omega-3 or omega-6    fatty acids.-   11. Subject had a significant history of drug/solvent abuse or had    positive test results at screening for DOA.-   12. Subject had a history of alcohol abuse, in the opinion of the    Investigator, or drank in excess of 28 units per week (males) or 21    units per week (females) at the time of screening.-   13. Subject was, in the opinion of the Investigator, not suitable    for participation in the study.-   14. Subject had participated in another clinical study with a study    drug/device within 3 months before the first day of administration    of study drug.-   15. Subject had a positive test result for HIV antibodies, Hepatitis    B surface antigen, or Hepatitis C antibodies at screening.-   16. Subject had a serious adverse reaction or significant    hypersensitivity to any drug.-   17. Subject had donated blood or blood products within 3 months    before screening.-   18. Subject had known hypersensitivity to any ingredients of the    study drug.

Subjects were free to withdraw consent from the study at any time forany reason. In addition, the Investigator could withdraw a subject fromstudy participation if, in the Investigator's opinion, it was in thebest interest of the subject. A subject would be withdrawn from thestudy for any of the following reasons:

-   -   Withdrawal of consent any time    -   Deviations from the protocol    -   Incidental illness    -   An AE (adverse effect).

Although a subject was not obliged to give a reason for prematurewithdrawal, the Investigator was to make a reasonable effort to obtainthe reason while fully respecting the subject's rights. If there was amedical reason for withdrawal of consent, the subject was to remainunder the supervision of the Investigator until the subject was insatisfactory health; the Investigator was to conduct a follow-upassessment.

If the Investigator deemed it was considered in the best interest of thesubject's welfare, the Investigator was to inform the subject's GeneralPractitioner of the medical reason for the subject's withdrawal from thestudy. Every effort was to be made to contact a subject who failed toreturn to the site for any follow-up appointments in order to ensurethat the subject was in satisfactory health.

In Part one of the study, subjects in Cohorts one to three wereadministered a single dose of study drug (either 500-, 1000-, or 2000-mgas 500-mg DS107G capsules or matching placebo capsules) in parallel onstudy Day 1 after at least an 8-hour fast, according to Table 15. Cohort4 was initiated following the review of safety data from Cohort 3 by theSMC.

Food effect was evaluated in Cohort two at least 14 days after the firstdose at which time a second single dose was administered. Subjects wereadministered a 1000-mg dose of study drug with 240 mL of water after a10-hour fasting period and 30 minutes after starting consumption of ameal. Subjects then refrained from food intake for at least 4 hoursafter dosing. Food consumption was standardized for at least 12 hoursafter dosing using a standardized high-fat meal (800 to 1000 kcal with500 to 600 kcal from fat and 250 kcal from carbohydrates). A typicalstandard test meal consisted of two eggs fried in butter, two strips ofbacon, two slices of toast with butter, 120 mL of hash brown potatoes,and 240 mL of whole milk.

In Part two of the study (Cohorts five and six), subjects wereadministered study drug (500-mg DS107G capsules or matching placebocapsules) once daily in the fasted state for 28 days. Subjects in Cohortfive received study drug first and, if they tolerated the 2000-mg dailydose for the first 14 days, subjects in Cohort six were started on the4000-mg daily dose for 28 days.

TABLE 15 Treatment Cohorts Number of Fasted or Cohort Subjects Dose (mg)capsules Fed State Part 1: Single Ascending Doses 1 8 500-mg DGLA orplacebo 1 Fasted capsules 2 8 1000-mg DGLA or placebo 2 Fasted capsules8 1000-mg DGLA or placebo 2 Fed capsules 3 8 2000-mg DGLA or placebo 4Fasted capsules 4 8 4000-mg DGLA or placebo 8 Fasted capsules Part 2:Multiple Ascending Doses for 28 Consecutive Days 5 8 2000-mg DGLA orplacebo 4 Fasted capsules 6 8 4000-mg DGLA or placebo 8 Fasted capsulesNote: Each DS107G capsule contained 500 mg DGLA.

Any escalation in the dose level or commencement of a subsequent cohortwas decided by the Principal Investigator. A minimum of 5 subjects withevaluable safety data from Cohort three (2000-mg dose) was requiredbefore commencing Cohort four (4000-mg dose). In Part 2, a minimum of 5subjects with 14 days of evaluable safety data from Cohort 5 (2000mg/day) was required before commencing Cohort six (4000 mg/day).

One strength of DS107G DGLA capsule was developed containing 500 mg ofDGLA free fatty acid (FFA). Capsules included the following excipients:DGLA FFA (stabilized with a nominal 2000 ppm dl-alpha tocopherol). Allexcipients in the capsule shell were commonly used in soft gelatinproducts and include a transmissible spongiform encephalopathy (TSE)certified gelatin shell containing the following ingredients: purifiedwater, the plasticizer glycerol, the colorant titanium dioxide, and theprocessing aids lecithin and medium chain triglyceride. The placebocapsule (DS107G Placebo capsule) for clinical studies consisted ofliquid paraffin encapsulated in a soft gelatin shell and was identicalin appearance to the DGLA capsule.

Subjects meeting the eligibility criteria were randomly assigned toreceive DGLA (500, 1000, 2000, 4000 mg doses) or matching placebocapsules using a randomization schedule. The randomization was blockrandomization with an active treatment to placebo ratio of 3:1. Therandomization schedule was generated by Planimeter using SAS® 9.1.3 SP4.

Subjects were not permitted to use prescribed medication during the 2weeks prior to dosing or to use over-the-counter preparations (includingvitamins and supplements) and hormonal contraceptives for 1 week beforedosing, with the exception of paracetamol, which was allowed up to 48hours before dosing. In addition, subjects were not allowed to usedietary supplements rich in omega-3 or omega-6 fatty acids. Subjectswere not permitted to consume alcohol in excess of 28 units per week(male subjects) or 21 units per week (female subjects).

Subjects were advised to avoid consuming food supplements rich inomega-3 or omega-6 fatty acids (e.g., cod liver oil capsules) for 4weeks prior to Screening and during the study.

Subjects were advised to avoid eating poppy seeds and food containingpoppy seeds for at least 24 hours before urine sample collection fortesting DOA as poppy seeds can sometimes cause a positive test result.

Subjects in Cohort two undergoing evaluation of the food effect on oral1000 mg DGLA capsules were restricted from food consumption for at least4 hours after dosing. Food consumption was standardized for at least 12hours post dose using a standardized high-fat meal.

Subjects were to refrain from taking any systemic or over-the-countermedication (including vitamins and supplements) during the study withthe exception of hormonal contraceptives. Paracetamol (at a dose up to 4g/day) was allowed up to 48 hours before dosing. Subjects were also torefrain from alcohol consumption from 48 hours prior to the firstadministration of study drug (Day 1) until the follow-up visit.

There were no restrictions on caffeine intake or tobacco use prior to orduring the study.

Subjects were required to avoid exercise and strenuous physical activityfor at least 3 to 4 hours before the blood was collected for theclinical laboratory test.

Analysis of plasma concentrations of DGLA (free and total) and free15-HETrE were performed in blood samples obtained from the subjects. Inaddition, skin blister fluids were obtained at days 1, 7, 14, and 28 foranalysis of free and total DGLA.

Plasma concentrations of dihydroxytestosterone (DHT) were assessed inPart 2 as a biomarker or exploratory efficacy endpoint.

Analysis of all plasma and skin blister fluid samples was performedusing validated methods. Concentrations of free and total DGLA weremeasured in plasma and skin blister fluid by liquid chromatography withtandem mass spectrometry (LC/MS/MS); the quantitation range was 100 to10,000 ng/mL for free DGLA and 5000 to 500,000 ng/mL for total DGLA.Plasma concentrations of free 15-HETrE were measured by LC/MS/MS with aquantitation range of 100 to 10,000 ng/mL. Plasma concentrations of DHTwere measured by LC/MS with a quantitation range of 0.02 to 1.5 ng/mL.

Subjects were monitored throughout confinement at the Phase 1 unit foradverse reactions to the study drug and/or procedures.

The pharmacokinetic assessments used in this study were standard forevaluation of potential therapeutic agents. The safety assessmentsincluded methods that were considered to be standard for a Phase 1clinical study.

The study was exploratory, and no formal power calculations wereperformed. The number of subjects planned for each cohort (8 subjects)was considered sufficient to allow assessment of the safety and systemicexposure of DS107G capsule.

Analysis of populations included the following:

-   -   The Intention-to-Treat (ITT) population consists of all        randomized subjects who had been administered at least 1 dose of        study drug.    -   The Per Protocol (PP) population consists of all subjects in the        ITT population who had no major protocol violation, as defined        in the SAP.    -   The PK population contains all subjects included in the PP        population who had evaluable PK data derived from plasma. A        plasma concentration observation was considered a valid,        evaluable measurement if the following data were available:        study identification number, randomization number, date and time        of sampling, dose and concentration. A series of such        measurements from the same sample were considered complete if        each protocol prescribed concentration was evaluable. Plasma PK        data were evaluable by definition if the data contained a        complete series of observations. Any missing plasma PK        observation would result in incomplete PK data, thus a subject        with any missing plasma PK observation was excluded from the PK        population. Subjects randomized to placebo were also excluded        from the PK population.

The populations defined above were generated separately for study Part 1and Part 2 data. Safety analysis was performed on the ITT population.

Safety analyses were performed on the safety population tabulated bytreatment arm, cohort, and visit. All safety data were characterized bydescriptive statistical tools. No hypothesis was set to investigate inthe study protocol. Evaluation of safety assessments was performed in adescriptive manner. Continuous variables were characterized by theirmean, standard deviation (SD), median, minimum and maximum values;discrete variables were characterized by their absolute (frequency) andrelative (percentage [%]) distribution. Treatment groups were also

Primary endpoints (PK characteristics derived from single and multipleoral doses of DS107G capsule) were derived with the help ofnon-compartmental PK modeling. Secondary endpoints consisted ofcharacterizing the effect of food on the PK of a single oral dose ofDS107G capsule and characterizing the PK of DGLA in human skin followingmultiple oral doses of DS107G capsule).

The secondary endpoints were reported using the following descriptiveanalytical tools: number of valid observations, mean, standard deviation(SD), median, minimum, and maximum values derived for continuousparameters grouped by treatment arm, visit and cohort.

No formal hypothesis testing was performed during evaluation of studydata. In Part 1 (single ascending dose), the extent of exposure was 1day by definition (in case of successful administration of the studydrug). In Part 2 (multiple ascending doses), the extent of exposure wascalculated as the day of last study drug intake minus the day of firststudy drug intake+1 (when no interruption in study drug administrationwas documented). In case of interruption(s) the result of the aboveformula was decreased with the number of interruption(s).

Statistical analysis of the PK data was performed using SAS Software(Version 9.1.3). No pharmacodynamics analysis was performed.

Plasma PK parameters of DGLA and of 15-HETrE were estimated usingmodel-independent techniques (non-compartmental analysis) and included:Cmax, tmax, Clast, Tlast, AUC0-24, AUC0-inf, AUClast, λz, CL, V, andt1/2 for Parts 1 and 2, and tmin, cmin, CLss, Vss, Cavg, and % PTF foronly Part 2 (steady-state) data.

All PK parameters were summarized descriptively, no formal statisticaltesting was performed due to the exploratory nature of the study.

In the single-dose setting (Part 1), study drug discontinuation was nota potential outcome in Cohorts one, three and four; therefore, noreporting activity was required for these cohorts. For Cohort 2, theenrolled subjects received study drug twice: a single dose administeredin the fasted state and a second single dose administered in the fedstate.

The following amendments were also made to the protocol.

-   -   Table of Clinical Studies added, to clarify previous clinical        studies with oral DGLA and their safety conclusions.    -   Risk/benefit assessment added, to clarify the assessed risks and        benefits of the proposed trial for the benefit of the        Investigator and Ethics Committee.    -   Enrollment criteria were changed, such that subjects who were        enrolled in Part 1 of the study (single-dose cohorts) could be        brought back and re-enrolled in Part 2 (multiple-dose cohorts),        provided they had no AEs related to study drug and they had a        washout period of at least 14 days before starting the        multiple-dose regimen. The rationale for this change was to aid        recruitment without compromising the safety of the volunteers.    -   Clinical laboratory tests for coagulation (prothrombin time and        activated partial prothrombin time [APTT]) were added as        assessments at all clinical laboratory assessment timepoints in        Part 2 of the study (multiple-dose Cohorts 5 and 6). These        assessments were added to monitor any potential changes in        clotting factors as an exploratory biomarker for future studies.    -   Ambiguous wording was replaced to clarify that single ECG        recordings would be obtained.    -   15-HETrE was removed as an analyte for testing in Part 2        (multiple-dose cohorts), as preliminary PK data from Part 1        cohorts revealed that all samples for 15-HETrE were below the        limit of quantification (BLQ).    -   In addition to free (unesterified) DGLA, the quantification of        “total” DGLA was added to all analyses, as the plasma PK profile        of the “total” may differ from the “free.”    -   Also, changes to the planned analyses were included to provide        additional analyses and statistical output.

RESULTS Single Dose Study.

Forty subjects were screened; of those 40, 4 subjects were excluded fornot fulfilling inclusion/exclusion requirements, and 4 subjects withdrewconsent. Disposition of the 32 subjects randomized to study drug ispresented in Table 16.

TABLE 16 Summary of Subject Disposition - ITT population (Single Dose).Dose Level of DS107G 500 mg 1000 mg 2000 mg 4000 mg Placebo¹ Overall (N= 6) (N = 6) (N = 6) (N = 6) (N = 8) (N = 32) n (%) n (%) n (%) n (%) n(%) n (%) Total number of subjects 6 (100) 6 (100) 6 (100) 6 (100) 8(100) 32 (100) Randomized 6 (100) 6 (100) 6 (100) 6 (100) 8 (100) 32(100) Treated 6 (100) 6 (100) 6 (100) 6 (100) 8 (100) 32 (100) Completed6 (100) 6 (100) 6 (100) 6 (100) 8 (100) 32 (100) Prematurediscontinuation 0 0 0 0 0  0 ITT = intent-to-treat ¹Of the 8 subjectswho received placebo, 2 subjects were randomized to each of the DS107Gdosing cohorts.

Plasma concentrations of free and total DGLA at baseline, beforeadministration of DS107G, were summarized in Table 17. Theseconcentrations were generally highly variable.

TABLE 17 Baseline Plasma Concentrations of DGLA (Single Dose,Pharmacokinetic Population) Dose Level of DS107G 500 mg 1000 mg 2000 mg4000 mg Analyte Fasted Fasted Fed Fasted Fasted (unit) (n = 5) (n = 6)(n = 6) (n = 6) (n = 6) Free DGLA Mean 237.0 302.3 262.3 172.8 378.2(ng/mL) (SD) (76.95) (29.80) (70.22) (29.67) (101.18) Min, Max 150, 357268, 347 133, 327 134, 204 241, 538 Total Mean 37740.0 40583.3 42550.033083.3 29900.0 DGLA (SD) (14371.60) (13664.47) (16206.02) (8247.52)(10164.05) (ng/mL) Min, Max 22000, 57100 26700, 63900 28400, 6660022700, 43200 19100, 45500 DGLA = dihomo-gamma-linolenic acid; Max =maximum; mean = arithmetic mean; min = minimum; N = number of subjectsproviding data; SD = standard deviation

Mean plasma concentrations after a single dose of DS107G are showngraphically by dose cohort for free DGLA in FIG. 6 (linear plot) andFIG. 7 (log-linear plot), and for total DGLA in FIG. 8 (linear plot) andFIG. 9 (log-linear plot).

After a single dose of DS107G capsules (500, 1000, 2000, and 4000 mg),inter-subject variability (measured by SD) of free and total DGLA washigh for both plasma concentrations and baseline-corrected PKparameters. Under fasted conditions, mean baseline-corrected C_(max) andAUC0-24 for both free and total DGLA increased in a linear manner (Table18, Table 19). The median time to maximum concentration (T_(max)) was 4hour for free DGLA (Table 18) and not consistent across the dose, withvalues of 8 hours at the 2 lower doses and 18 hours at the 2 higherdoses (Table 19). Although baseline-corrected elimination PK parameterscould be determined for less than half the subjects in some cohorts,there was no evidence of nonlinear pharmacokinetics in eliminationhalf-life or clearance for either free or total DGLA (Table 19 and Table20). Although not evaluated statistically, administration of a single1000-mg dose of DS107G under fasted conditions resulted in anapproximately 50% higher rate and extent of total DGLA absorption basedon baseline corrected mean Cmax and AUC0-24 (Table 19).

TABLE 18 Baseline-corrected Plasma Pharmacokinetic Parameters for FreeDGLA after Fasted Administration (Single Dose, PharmacokineticPopulation) Dose Level of DS107G Parameter 500 mg 1000 mg 2000 mg 4000mg (unit) Statistic (n = 5) (n = 6) (n = 6) (n = 6) t_(max) N 5 6 6 6(h) Median 4.00 4.00 4.00 4.00 Min, Max 4.0, 4.0 4.0, 4.0 4.0, 8.0 4.0,6.0 C_(max) N 5 6 6 6 (ng/mL) Mean 934.6 1602.8 2074.0 3021.8 SD 776.871202.43 1276.03 1581.97 C_(max)/Dose N 5 6 6 6 (1/kL) Mean 1.869 1.6031.037 0.755 SD 1.5537 1.2024 0.6380 0.3955 AUC₀₋₂₄ N 5 6 6 6 (ng · h/mL)Mean 2728.1 3998.5 6429.0 9436.8 SD 2253.49 1885.27 2119.39 2227.25AUC₀₋₂₄/Dose N 5 6 6 6 (h/kL) Mean 5.46 4.00 3.21 2.36 SD 4.507 1.8851.060 0.557 λ_(z) N 2 5 5 5 (1/h) Mean 0.0035 0.0487 0.0200 0.0603 SD0.00101 0.05133 0.02445 0.10642 t_(1/2) N 2 5 5 5 (h) Median 206.3328.30 57.33 67.53 Min, Max 164.3, 248.4   6.2, 3226.3  11.0, 181.1  2.8,80.9 AUC_(0-inf) N 2 5 5 5 (ng · h/mL) Mean 21483.3 218955.6 26877.017113.6 SD 14902.37 474291.75 19394.67 3545.72 AUC_(0-inf)/Dose N 2 5 55 (h/kL) Mean 42.97 218.96 13.44 4.28 SD 29.805 474.292 9.697 0.886 CL/FN 2 5 5 5 (kL/h) Mean 0.0306 0.1866 0.1339 0.2420 SD 0.02126 0.181540.11952 0.05066 Vz/F N 2 5 5 5 (kL) Mean 10.035 7.356 8.977 17.460 SD8.9574 9.1148 4.0610 11.5516 Max = maximum; mean = arithmetic mean; min= minimum; N = number of subjects providing data; SD = standarddeviation Note: Predose DGLA concentration was subtracted fromsubsequent concentrations before parameter calculation; negative valueswere replaced by zero.

TABLE 19 Baseline-corrected Plasma Pharmacokinetic Parameters for TotalDGLA after Fasted Administration (Single Dose, PharmacokineticPopulation) Dose Level of DS107G Parameter 500 mg 1000 mg 2000 mg 4000mg (unit) (n = 5) (n = 6) (n = 6) (n = 6) t_(max) N 5 6 6 6 (h) Median8.00 8.00 18.00 18.00 Min, Max 6.0, 312.0 0.0, 312.0 6.0, 24.0  8.0,24.0  C_(max) N 5 6 6 6 (ng/mL) Mean 7200.0 15750.0 17733.3 25883.3 SD2801.79 16326.02 9867.66 13658.32 C_(max)/Dose N 5 6 6 6 (1/kL) Mean14.400 15.750 8.867 6.471 SD 5.6036 16.3260 4.9338 3.4146 AUC₀₋₂₄ N 5 46 6 (ng · h/mL) Mean 39556.6 215369.7 210508.9 437754.0 SD 37969.05346249.37 167660.31 278838.30 AUC₀₋₂₄/Dose N 5 4 6 6 (h/kL) Mean 79.11215.37 105.25 109.44 SD 75.938 346.249 83.830 69.710 λ_(z) N 2 2 2 5(1/h) Mean 0.0013 0.0136 0.0084 0.0102 SD 0.00042 0.01381 0.003010.00993 t_(1/2) N 2 2 2 5 (h) Median 552.07 105.42 88.04 71.03 Min, Max428.3, 675.9  29.7, 181.2  65.8, 110.3  26.7, 464.7  AUC_(0-inf) N 2 2 25 (ng · h/mL) Mean 4415966.6 4031410.9 2276495.1 5973518.8 SD 723917.025304874.41 1948487.62 7261476.90 AUC_(0-inf)/Dose N 2 2 2 5 (h/kL) Mean8831.93 4031.41 1138.25 1493.38 SD 1447.834 5304.874 974.244 1815.369CL/F N 2 2 2 5 (kL/h) Mean 0.0001 0.0018 0.0014 0.0036 SD 0.000020.00243 0.00119 0.00519 Vz/F N 2 2 2 5 (kL) Mean 0.089 0.093 0.203 0.275SD 0.0140 0.0843 0.2136 0.1569 Max = maximum; mean = arithmetic mean;min = minimum; N = number of subjects providing data; SD = standarddeviation Note: Predose DGLA concentration was subtracted fromsubsequent concentrations before parameter calculation; negative valueswere replaced by zero.

The effect of food on the single-dose baseline-corrected PK of free andtotal DGLA using the 1000-mg dose (Cohort two) was evaluated andreported in Table 21. Briefly, the mean baseline-corrected free DGLACmax was about 3 fold higher and occurred 1 hour (median) sooner underfasted conditions than under fed conditions (Table 21). Mean free DGLAbaseline-corrected AUC0-24 under fasted conditions was about 2 foldhigher than the value under fed conditions. Thus an increased rate andextent of DGLA absorption under fasted conditions was observed. Therewere no clear differences for fasted vs fed conditions in elimination offree DGLA (Table 20).

For total DGLA, mean baseline-corrected Cmax was about 1.5-fold higherand tmax occurred about 50% sooner (median, 8 vs 15 h) under fastedconditions than under fed conditions (Table 20). Mean baseline-correctedAUC0-24 under fasted conditions was about 1.8-fold higher than under fedconditions. Just under half of the subjects (2/6 fasted, 3/6 fed) hadsufficient data to estimate total DGLA λz, t1/2, clearance, and volumeof distribution. These data suggest an increased rate and extend oftotal DGLA absorption under fasted condition. No reliable conclusionregarding total DGLA elimination or volume distribution due to smalldata population.

Based on the PK parameters shown in Table 20, mean baseline-correctedCmax was ˜10 fold (fasted) and ˜20-fold (fed) higher for total DGLA thanfor free DGLA. Mean baseline-corrected AUC0-24 was ˜54-fold (fasted) and˜56-fold (fed) higher for total DGLA than for free DGLA.

TABLE 20 Baseline-corrected Plasma Pharmacokinetic Parameters for DGLA,After Fasted vs Fed Administration (Single Dose, PharmacokineticPopulation) 1000 mg DS107G Free DGLA Total DGLA Parameter Fasted FedFasted Fed (unit) Statistic (n = 6) (n = 6) (n = 6) (n = 6) t_(max) N 66 6 6 (h) Median 4.00 5.00 8.00 15.00 Min, Max 4.0, 4.0 4.0, 8.0 0.0,312.0 6.0, 24.0 C_(max) N 6 6 6 6 (ng/mL) Mean 1602.8 520.2 15750.010316.7 SD 1202.43 235.54 16326.02 2043.93 C_(max)/Dose N 6 6 6 6 (1/kL)Mean 1.603 0.520 15.750 10.317 SD 1.2024 0.2355 16.3260 2.0439 AUC₀₋₂₄ N6 6 4 6 (ng · h/mL) Mean 3998.5 2102.7 215369.7 117389.7 SD 1885.271174.23 346249.37 70263.85 AUC₀₋₂₄/Dose N 6 6 4 6 (h/kL) Mean 4.00 2.10215.37 117.39 SD 1.885 1.174 346.249 70.264 λ_(z) N 5 3 2 3 (1/h) Mean0.0487 0.0403 0.0136 0.0805 SD 0.05133 0.06154 0.01381 0.06155 t_(1/2) N5 3 2 3 (h) Median 28.30 140.86 105.42 6.62 Min, Max   6.2, 3226.3  6.2,149.4 29.7, 181.2  5.5, 66.0 AUC_(0-inf) N 5 3 2 3 (ng · h/mL) Mean218955.6 29150.8 4031410.9 267053.2 SD 474291.75 23323.42 5304874.41335666.87 AUC_(0-inf)/Dose N 5 3 2 3 (h/kL) Mean 218.96 29.15 4031.41267.05 SD 474.292 23.323 5304.874 335.667 CL/F N 5 3 2 3 (kL/h) Mean0.1866 0.1658 0.0018 0.0096 SD 0.18154 0.24651 0.00243 0.00702 Vz/F N 53 2 3 (kL) Mean 7.356 4.623 0.093 0.128 SD 9.1148 0.5166 0.0843 0.0209Max = maximum; mean = aritnmetic mean; min = minimum; N = number ofsubjects providing data; SD = standard deviation Note: Predose DGLAconcentration was subtracted from subsequent concentrations beforeparameter calculation; negative values were replaced by zero.

DSI07G was well tolerated as single dose at amounts 500, 1000, 2000, or4000 mg to healthy volunteers. The most common TEAE reported were mildto moderate diarrhea (reported term: loose stool), by a similarpercentage of subjects between the active-treatment and placebo-controlsubjects (incidence: 5/24 [20.8%] active-treated subjects vs 2/8 [25.0%]placebo-control subjects). The diarrhea events were of relatively shortduration, and all (including those occurring in placebo-controlsubjects) were considered by the Investigator to be possibly related tostudy drug. Of note, subjects who received a second single dose ofDS107G and who had TEAEs of diarrhea in the fasted state did not have arecurrence of diarrhea All other TEAEs occurred in only 1 subject eachand only in the active-treatment groups, including: mild infection,oropharyngeal pain, and pharyngitis; and moderately severe pyrexia andurinary tract infection after dosing in the fed state.

Multiple-Dose Results—Study Part 2

Mean plasma concentrations and the average concentration at steady-statefor Part 2 are shown graphically by dose cohort for free DGLA in FIG. 10(linear plot) and FIG. 11 (log-linear plot), and for total DGLA in FIG.12 (linear plot) and FIG. 13 (log-linear plot). On Days 1 and 28, meanconcentrations of free DGLA peaked at about 4 hours after dosing,whereas no mean peak concentration was evident for total DGLA. Meanconcentrations of both free and total DGLA increased over time withrepeated dosing, though the increase for total DGLA was more pronounced.Plasma concentrations appeared to reach steady-state by around Day 14for both doses (2000 and 4000 mg daily) and analytes (free and totalDGLA) based on visual inspection of the mean concentration plots. Whenthe dose doubled from 2000 to 4000 mg daily, the average concentrationat steady state increased 1.6-fold for free DGLA but only 1.2-fold fortotal DGLA, suggesting one or more saturable processes at the higherdose.

PK parameter was computed after correcting the dosed DGLA concentrationswith baseline DGLA concentrations.

The plasma baseline corrected pharmacokinetics for free DGLA is reportedin Table 21. Briefly, mean free DGLA baseline-corrected C_(max) and AUCwere higher in the higher DS107G dose cohort on both days evaluated.Mean baseline-corrected C_(max) for the 4000 mg dose was ˜3-fold higherthan for the 2000 mg dose on Day 1 but only ˜1.4 fold higher on Day 28.Mean baseline-corrected AUC0-24 for the 4000 mg dose was ˜2.5 foldhigher than for the 2000 mg dose on Day 1 and only ˜1.7-fold higher onDay 28. The changes with dose were linear for baseline-corrected Cmaxand AUC0-24 on Day 1, but only for baseline-corrected AUC0-24 on Day 28.High inter-subject variability might have caused this inconsistency.Median t_(max) was similar for both dose cohorts on Day 1 and Day 28,with values of 4 or 5 hours. Median elimination t1/2 was longer on Day28 than on Day 1, with the value on Day 28 dependent on the timeinterval evaluated. Mean clearance decreased and mean volume ofdistribution increased with multiple doses.

TABLE 21 Plasma Baseline-corrected Pharmacokinetic Parameters for FreeDGLA (Multiple-dose, Pharmacokinetic Population) Dose Level of DS107GDay 1 Day 28 Day 28 0 to 24 hours 0 to 24 hours 0 to 168 hours Parameter2000 mg 4000 mg 2000 mg 4000 mg 2000 mg 4000 mg (unit) Statistic (n = 6)(n = 6) (n = 6) (n = 6) (n = 6) (n = 6) t_(max) N 6 6 6 6 6 6 (h) Median4.00 5.00 4.00 4.00 4.00 4.00 Min, Max 4.0, 6.0 2.0, 8.0  2.0, 4.0  4.0,4.0  2.0, 4.0  4.0, 4.0  C_(max) N 6 6 6 6 6 6 (ng/mL) Mean 850.5 2641.51999.3 2873.2 1999.3 2873.2 SD 458.84 1734.46 1003.19 1246.97 1003.191246.97 C_(max)/Dose N 6 6 6 6 6 6 (1/kL) Mean 0.425 0.660 1.000 0.7181.000 0.718 SD 0.2294 0.4336 0.5016 0.3117 0.5016 0.3117 AUC₀₋₂₄ N 6 6 66 6 6 (ng · h/mL) Mean 4548.7 11441.4 9248.1 16155.2 9248.1 16155.2 SD3330.51 5313.29 2559.31 6367.97 2559.31 6367.97 AUC₀₋₂₄/Dose N 6 6 6 6 66 (h/kL) Mean 2.27 2.86 4.62 4.04 4.62 4.04 SD 1.665 1.328 1.280 1.5921.280 1.592 λ_(z) N 2 6 3 5 5 6 (1/h) Mean 0.2133 0.1504 0.1282 0.05960.0250 0.0404 SD 0.16812 0.10843 0.15619 0.02019 0.02983 0.05960 t_(1/2)N 2 6 3 5 5 6 (h) Median 4.71 5.92 14.04 12.72 63.59 48.18 Min, Max 2.1,7.3 2.0, 18.3 2.2, 25.6 7.5, 16.5 9.0, 132.1 4.3, 65.7 AUC_(0-inf) N 2 63 5 5 6 (ng · h/mL) Mean 5955.6 13900.3 10389.0 22926.3 43024.8 43184.3SD 5216.07 4993.97 2677.43 6485.37 35641.74 21443.83 AUC_(0-inf)/Dose N2 6 3 5 5 6 (h/kL) Mean 2.98 3.48 5.19 5.73 21.51 10.80 SD 2.608 1.2481.339 1.621 17.821 5.361 CL/F N 2 6 3 5 5 6 (kL/h) Mean 0.5448 0.33250.2002 0.1865 0.0949 0.1266 SD 0.47711 0.16310 0.04493 0.05432 0.086360.08671 Vz/F N 2 6 3 5 5 6 (kL) Mean 2.426 3.063 3.592 3.493 4.470 5.273SD 0.3245 1.9716 2.5139 1.5255 1.5179 2.1568 Max = maximum; mean =arithmetic mean; min = minimum; N = number of subjects providing data;SD = standard deviation Note: Predose DGLA concentration was subtractedfrom subsequent concentrations before parameter calculation; negativevalues were replaced by zero.

The steady state plasma baseline corrected pharmacokinetics for freeDGLA is reported in Table 22. Briefly, the plasma concentrations of freeand total DGLA increased with repeated dosing, and achieved steady-stateat approximately Day 14. When at steady state, evaluated on Day 28 (0-24hours), Peak Trough Fluctuation (PTF) was quite high for both dosecohorts (mean, ˜430% and ˜490%; Table 9). The mean accumulation ratio(AR) was greater for the 2000 mg than the 4000 mg dose cohort for bothCmax and AUC (ARs of ˜2.8 and ˜3.3 for 2000 mg vs ˜1.4 and ˜1.6 for 4000mg. The data suggest saturable kinetics and/or presence of change in thedistribution volume with repeated dosing of free DGLA.

TABLE 22 Steady-state Plasma Baseline-corrected PharmacokineticParameters for Free DGLA (Multiple-dose, Pharmacokinetic Population) Day28: 0 to 24 hours Dose Level of DS107G Parameter 2000 mg 4000 mg (unit)(n = 6) (n = 6) t_(min) N 6 6 (h) Median 8.00 8.00 Min, Max 6.0, 24.00.0, 12.0 C_(min) N 6 6 (ng/mL) Mean 100.8 147.0 SD 103.34 230.37CL_(ss)/F N 6 6 (kL/h) Mean 0.2295 0.2858 SD 0.05874 0.12493 Vz_(ss)/F N3 5 (kL) Mean 5.3096 5.7563 SD 4.02532 3.12495 C_(avg) N 6 6 (ng/mL)Mean 385.3 673.1 SD 106.64 265.33 % PTF N 6 6 (%) Mean 492.54 431.97 SD212.957 207.312 AR(C_(max)) N 6 6 Mean 2.796 1.385 SD 1.7385 0.7726AR(AUC) N 6 6 Mean 3.340 1.561 SD 2.4958 0.6377 AR = accumulation ratio;Max = maximum; mean = arithmetic mean; min = minimum; N = number ofsubjects providing data; PTF = peak trough fluctuation; SD = standarddeviation Note: Predose DGLA concentration was subtracted fromsubsequent concentrations before parameter calculation; negative valueswere replaced by zero.

The plasma baseline corrected pharmacokinetics for total DGLA isreported in Table 23. Briefly, mean total DGLA baseline-correctedC_(max) and AUC0-24 were higher in the higher DS107G dose cohort on bothdays evaluated, as expected. Mean baseline-corrected C_(max) and AUC0-24for the 4000 mg dose were ˜1.5- and ˜1.5-fold higher, respectively, thanfor the 2000 mg dose on Day 1 but only ˜1.2- and ˜1.4-fold higher thanfor the 2000 mg dose on Day 28.

The changes with dose in baseline-corrected C_(max) and AUC4 were notlinear for total DGLA on either day evaluated. High inter-subjectvariability might have caused this inconsistency. Median Tmax occurredsooner with multiple doses (8-10 h) than with a single dose (10 18 h) inboth dose cohorts. Median total DGLA elimination t1/2 for the 2000 mgdose cohort was 34.4 to 44.0 hours when assessed over 24 hours (Days 1and 28, respectively), and 62.6 hours on Day 28 when evaluated over 0 to168 hours. Mean clearance and volume of distribution decreased withmultiple doses.

TABLE 23 Plasma Baseline-corrected Pharmacokinetic Parameters for TotalDGLA (Multiple-dose, Pharmacokinetic Population) Dose Level of DS107GDay 1 Day 28 Day 28 0 to 24 hours 0 to 24 hours 0 to 168 hours Parameter2000 mg 4000 mg 2000 mg 4000 mg 2000 mg 4000 mg (unit) Statistic (n = 6)(n = 6) (n = 6) (n = 6) (n = 6) (n = 6) t_(max) N 6 6 6 6 6 6 (h) Median10.00 18.00 8.00 10.00 8.00 10.0 Min, Max 8.0, 12.0 12.0, 24.0  6.0,24.0 8.0, 24.0  6.0, 24.0  8.0, 24.0 C_(max) N 6 6 6 6 6 6 (ng/mL) Mean18100.0 27866.7 75583.3 90866.7 75583.3 90866.7 SD 15113.44 10391.6630385.55 33000.16 30385.55 33000.16 C_(max)/Dose N 6 6 6 6 6 6 (1/kL)Mean 9.050 6.967 37.792 22.717 37.792 22.717 SD 7.5567 2.5979 15.19288.2500 15.1928 8.2500 AUC₀₋₂₄ N 6 6 6 6 6 6 (ng · h/mL) Mean 298184.2456199.5 1351372.5 1860574.3 1351372.5 1860574.3 SD 336969.41 151586.31379053.12 760386.93 379053.12 760386.93 AUC₀₋₂₄/Dose N 6 6 6 6 6 6(h/kL) Mean 149.09 114.05 675.69 465.14 675.69 465.14 SD 168.485 37.897189.527 190.097 189.527 190.097 λ_(z) N 3 0 4 3 6 6 (1/h) Mean 0.2210.0199 0.0471 0.0169 0.0194 SD 0.02180 0.00966 0.05409 0.01064 0.00927t_(1/2) N 3 0 4 3 6 6 (h) Median 44.04 34.40 36.57 62.64 39.21 Min, Max14.9, 162.5 22.2, 89.2 6.3, 53.8 20.0, 74.9 19.7, 70.9 AUC_(0-inf) N 3 04 3 6 6 (ng · h/mL) Mean 4161212.0 5159152.1 4779645.9 5027822.86303903.1 SD 6203131.92 4269186.29 2974257.92 2286762.41 4913744.09AUC_(0-inf)/Dose N 3 0 4 3 6 6 (h/kL) Mean 2080.61 2579.58 1194.912513.91 1575.98 SD 3101.566 2134.593 743.564 1143.381 1228.436 CL/F N 30 4 3 6 6 (kL/h) Mean 0.0027 0.0006 0.0012 0.0005 0.0009 SD 0.002660.00041 0.00099 0.00027 0.00056 Vz/F N 3 0 4 3 6 6 (kL) Mean 0.106 0.0290.035 0.032 0.046 SD 0.0597 0.0095 0.0114 0.0133 0.0133 Max = maximum;mean = arithmetic mean; min = minimum; N = number of subjects providingdata; SD = standard deviation Note: Predose DGLA concentration wassubtracted from subsequent concentrations before parameter calculation;negative values were replaced by zero.

The steady state plasma baseline corrected pharmacokinetics for freeDGLA is reported in Table 24. Briefly, at steady-state, evaluated on Day28 (0-24 hours), Peak Trough Fluctuation (PTF) was quite high for bothdose cohorts (mean, 62.5% and 44.9%). The mean AR was greater for the2000 mg than the 4000 mg dose cohort for both C_(max) and AUC. The datasuggest saturable kinetics and/or presence of change in the distributionvolume with repeated dosing of total DGLA.

TABLE 24 Steady-state Plasma Baseline-corrected PharmacokineticParameters for Total DGLA (Multiple-dose, Pharmacokinetic Population)Day 28: 0 to 24 hours Dose Level of DS107G Parameter 2000 mg 4000 mg(unit) Statistic (n = 6) (n = 6) t_(min) N 6 6 (h) Median 1.50 2.00 Min,Max 0.0, 6.0 2.0, 24.0 C_(min) N 6 6 (ng/mL) Mean 37933.3 59216.7 SD10890.12 37002.29 CL_(ss)/F N 6 6 (kL/h) Mean 0.0016 0.0025 SD 0.000510.00107 Vz_(ss)/F N 4 3 (kL) Mean 0.0980 0.1047 SD 0.04161 0.07339C_(avg) N 6 6 (ng/mL) Mean 56307.2 77523.9 SD 15793.88 31682.79 % PTF N6 6 (%) Mean 62.50 44.90 SD 31.403 38.967 AR(C_(max)) N 6 6 Mean 5.1563.440 SD 2.2542 1.2090 AR(AUC) N 6 6 Mean 7.991 4.163 SD 5.9537 1.2331

Mean free DGLA concentrations in skin blister fluid are shown by dosecohort in FIG. 14 (linear plot) and FIG. 15 (log-linear plot). Meanconcentrations approximately doubled with a doubling in dose (based onconcentrations from Days 1, 8, 14, and 28), and accumulated withrepeated doses in both regimens. Mean free DGLA concentrations on Day 28were about 3-fold higher than those on Day 1 for both 2000 and 4000 mgdaily.

Mean total DGLA concentrations in skin blister fluid are shown by dosecohort in FIG. 16 (linear plot) and FIG. 17 (log-linear plot). Meanconcentrations of total DGLA increased about 1.4-fold with a doubling indose (based on concentrations from Days 1, 8, 14, and 28). Mean totalDGLA concentrations on Day 28 were about 2.5- and 3-fold higher thanthose on Day 1 for 2000 and 4000 mg daily, respectively.

The concentration profiles in plasma and skin blister fluid wereoverlaid and mean concentrations of free DGLA were somewhat similar inplasma and skin blister fluid for the same DS107G dose at Day 8 and Day14 (but not Day 28). FIG. 18 [linear plot] and FIG. 19 [log-linearplot]), suggest that free DGLA distributes into plasma and skinsimilarly.

For total DGLA, mean concentrations were much higher in plasma than inskin blister fluid for the same DS107G dose after Day 1 (FIG. 20 [linearplot] and FIG. 21 [log-linear plot]), indicating that total DGLA isfound more readily in plasma than into skin. The mechanism for limiteddistribution of total DGLA into skin is most likely related to the lowerquantity of lipids in the skin compared with plasma.

Plasma dihydrotestosterone (DHT) concentrations were quantified as anexploratory efficacy endpoint or biomarker. Inter-subject variability inthe concentration data was high based on the SD at most time points.Mean plasma concentrations of DHT are shown by dose cohort in FIG. 22(linear) and FIG. 23 (log-linear).

None of the samples had measurable free 15-HETrE concentrations; allconcentrations were below the LLOQ (100 ng/mL).

There were no deaths for the multi-dose study. When administered tohealthy volunteers as 2000- or 4000-mg doses once daily for 28consecutive days, DS107G was tolerated well, with the worst TEAE beingmild to moderate diarrhea (reported term: loose stool) of relativelyshort duration. A majority of the events of diarrhea (incidence 7/12[43.8%] active-treated subjects vs 0/4 [0.0%] placebo-control subjects)were considered by the Investigator to be possibly related to studydrug. A higher proportion of subjects reported diarrhea in the 4000-mggroup (4/6 [66.7%]) than in the 2000-mg group (3/6 [50%]). The incidenceof TEAEs among active-treated subjects was much higher than that of theplacebo-control subjects (11/12 [91.73%] subjects reporting a total of52 TEAEs vs 1/4 [25.0%] subjects reporting a total of I TEAE,respectively). There were no severe TEAEs, and other than TEAEs ofdiarrhea, all events were considered to be either not or unlikelyrelated to study drug. Nausea was the next most commonly reported TEAE(10 events among 4/6 [66.7%] subjects in the 4000-mg treatment group);9/10 of the events of nausea were mild and the other was moderate inseverity. Other than diarrhea, all remaining TEAEs were reported in 2subjects each (bronchitis and nasopharyngitis) or 1 subject each(abdominal pain, asthenia, pyrexia, blood CPK increased, CRP increased,WBC count decreased, dizziness, headache, cough, and haematoma), themajority of which were considered by the Investigator to be unlikely ornot related to study drug. Other TEAEs considered to be possibly relatedto study drug were abdominal pain and asthenia (Reported Term“weakness”), both of which had temporal associations with events ofloose stool.

No clinically significant abnormalities in the vital sign or ECGobserved in any patients in multiple dose study.

1. A pharmaceutical composition comprising DGLA or a derivative thereofencapsulated in a capsule shell.
 2. The pharmaceutical composition ofclaim 1, wherein the capsule shell comprises gelatin and glycerol. 3.The pharmaceutical composition of claim 1, wherein up to about 1 g ofDGLA or a derivative thereof is encapsulated in the capsule shell. 4.The pharmaceutical composition of claim 1, wherein about 200 mg to about1 g of DGLA or a derivative thereof is encapsulated in the capsuleshell.
 5. The pharmaceutical composition of claim 1, wherein the gelatincomprises lime bone gelatin and the capsule shell further comprises oneor more of: glycerol, purified water, titanium dioxide, medium chaintriglycerides and/or lecithin.
 6. The pharmaceutical composition ofclaim 1, comprising about 200 mg to about 1 g of DGLA and wherein thecomposition comprises not less than 95% by weight DGLA, not more than 5%of any individual fatty acid selected from 20:2ω6, 20:3ω3, 20:4ω6,20:4ω3 and 20:5ω3, and not more than 2% by weight unidentified relatedsubstances.
 7. A method of treating disease or disorder in a subject inneed thereof, the method comprising orally administering to the subjecta pharmaceutical composition comprising DGLA or a derivative thereof. 8.The method of claim 7, wherein the pharmaceutical composition comprisesDGLA or a derivative thereof encapsulated in a capsule shell.
 9. Themethod of claim 8, wherein the composition is administered to thesubject in an amount sufficient to provide about 0.2 g to about 8 g ofDGLA per day.
 10. The method of claim 7 wherein the composition isadministered in 1 to 8 capsules per day.
 11. The method of claim 10wherein each capsule comprises about 200 mg to about 1 g of DGLA. 12.The method of claim 7, wherein the disease or disorder is selected from:skin disorders and diseases including acne vulgaris, acne rosacea,atopic dermatitis, psoriasis, pruritus/itch, radiation protection, dryskin, smooth skin, healthy skin, anti-aging, and photoprotection;urinary disorders and diseases including bladder cancer, cystocele,hematuria, interstitial cystitis, neurogenic bladder, Peyronie'sdisease, prostate disease, incontinence, urinary tract infection andvasicoureteral reflux; renal disease and disorders including kidneyfailure, acute kidney injury, chronic kidney disease, and polycystickidney disease; rheumatic disease including ankylosing spondylitis,fibromyalgia, gout, infectious arthritis, lupus, osteoarthritis,polymyalgia rheumatic, psoriatic arthritis, reactive arthritis,rheumatoid arthritis, sclerodoma; respiratory disorders includinginflammatory lung disease, respiratory tract infections, pleural cavitydisease, pulmonary vascular disease, pneumonia, pulmonary embolism, andlung cancer; and cardiovascular disorders including acute cardiacischemic events, acute myocardial infarction, angina, arrhythmia, atrialfibrulation, atherosclerosis, arterial fibrillation, cardiacinsufficiency, cardiovascular disease, chronic heart failure, chronicstable angina, congestive heart failure, coronary artery disease,coronary heart disease, deep vein thrombosis, diabetes, diabetesmellitus, diabetic neuropathy, diastolic dysfunction in subjects withdiabetes mellitus, edema, essential hypertension, eventual pulmonaryembolism, fatty liver disease, heart disease, heart failure, homozygousfamilial hypercholesterolemia (HoFH), homozygous familialsitosterolemia, hypercholesterolemia, hyperlipidemia, hypertension,hypertriglyceridemia, metabolic syndrome, mixed dyslipidemia, moderateto mild heart failure, myocardial infarction, obesity management,paroxysmal atrial/arterial fibrillation/fibrulation/flutter, paroxysmalsupraventricular tachycardias (PSVT), particularly severe or rapid onsetedema, platelet aggregation, primary hypercholesterolemia, primaryhyperlipidemia, pulmonary arterial hypertension, pulmonary hypertension,recurrent hemodynamically unstable ventricular tachycardia (VT),recurrent ventricular arrhythmias, recurrent ventricular fibrillation(VF), ruptured aneurysm, sitisterolemia, stroke, supraventriculartachycardia, symptomatic atrial fibrillation/flutter, tachycardia,type-II diabetes, vascular disease, venous thromboembolism, andventricular arrhythmias.
 13. A pharmaceutical composition comprisingDGLA, wherein upon oral administration of a single dose of saidcomposition comprising about 200 mg to about 8000 mg of DGLA to a fastedsubject the subject exhibits one or more of the following outcomes: (a)a free DGLA C_(max) of about 400 ng/ml to about 4500 ng/ml, about 500ng/ml to about 3400 ng/ml, about 600 ng/ml to about 3300 ng/ml, or about700 ng/ml to about 3200 ng/ml; (b) a free DGLA AUC₀₋₂₄ of about 1500ng·h/ml to about 12000 ng·h/ml, about 2000 ng·h/ml to about 11000ng·h/ml or about 2500 ng·h/ml to about 10000 ng·h/ml; (c) a free DGLAAUC₀₋₂₄/dose of about 1.5 to about 10 h/kL, about 1.7 to about 8 h/kL orabout 2 to about 6 h/kL; (d) a free DGLA t_(max) (h) of about 2 to about10 hours or about 3 to about 8 hours; (e) a total DGLA C_(max) of about4000 ng/ml to about 45000 ng/ml, about 5000 ng/ml to about 34000 ng/ml,about 6000 ng/ml to about 33000 ng/ml, or about 7000 ng/ml to about32000 ng/ml; (f) a total DGLA AUC₀₋₂₄ of about 15000 ng·h/ml to about900,000 ng·h/ml, about 20,000 ng·h/ml to about 500,000 ng·h/ml or about25,000 ng·h/ml to about 225,000 ng·h/ml; (g) a total DGLA AUC₀₋₂₄/doseof about 50 to about 400 h/kL, about 60 to about 250 h/kL or about 70 toabout 225 h/kL; (h) a total DGLA t_(max) (h) of about 2 to about 25hours or about 3 to about 20 hours; (i) a total DGLA C./dose of about 2(1/kL) to about 25 (1/kL), about 4 (1/kL) to about 20 (1/kL) or about 5(1/kL) to about 17 (1/kL); (j) a free DGLA C./dose of about 0.3 (1/kL)to about 2.5 (1/kL); (k) a ratio of total DGLA C_(max) to free DGLA C.of about 5:1 to about 12:1, about 6:1 to about 10:1 or about 7:1 toabout 10:1; and/or (l) a ratio of free DGLA plasma to DGLA skin asmeasured in skin blister fluid from about 0.2:1 to about 5:1, about0.5:1 to about 2.5:1 or about 0.6:1 to about 1.5:1.
 14. Thepharmaceutical composition of claim 13, wherein upon oral administrationof a single dose of said composition comprising about 200 mg to about8000 mg of DGLA to a fasted subject the subject exhibits one or more ofthe following outcomes: (a) a free DGLA C_(max) of about 700 ng/ml toabout 3200 ng/ml; (b) a free DGLA AUC₀₋₂₄ of about 2500 ng·h/ml to about10000 ng·h/ml; (c) a free DGLA AUC₀₋₂₄/dose of about 2 to about 6 h/kL;(d) a free DGLA t_(max)(h) of about 3 to about 8 hours; (e) a total DGLAC. of about 7000 ng/ml to about 32000 ng/ml; (f) a total DGLA AUC₀₋₂₄ ofabout 25,000 ng·h/ml to about 225,000 ng·h/ml; (g) a total DGLAAUC₀₋₂₄/dose 70 to about 225 h/kL; (h) a total DGLA t_(max) (h) of about3 to about 20 hours; (i) a total DGLA Cmax/dose of about 2 (1/kL) toabout 25 (1/kL), about 4 (1/kL) to about 20 (1/kL) or about 5 (1/kL) toabout 17 (1/kL); (j) a free DGLA Cmax/dose of about 0.3 (1/kL) to about2.5 (1/kL); (k) a ratio of total DGLA C. to free DGLA C_(max) of about7:1 to about 9:1; and/or (l) a ratio of free DGLA plasma to DGLA skin asmeasured in skin blister fluid from about 0.6:1 to about 1.5:1;
 15. Thepharmaceutical composition of claim 13, wherein upon oral administrationof multiple daily doses of said composition over a period of about 2 toabout 30 days, each daily dose comprising about 200 mg to about 8000 mgof DGLA, to a fasted subject the subject exhibits one or more of thefollowing outcomes: (a) a steady state free DGLA plasma level (C_(avg))of up to about 2000 ng/ml, up to about 750 ng/ml, or up to about 350ng/ml; (b) a steady state total DGLA plasma level (C_(avg)) of up to250,000 ng/ml, up to 180,000 ng/ml, up to 150,000 ng/ml, up to 125,000ng/ml or up to 100,000 ng/ml; and/or (c) a ratio of free DGLA plasma toDGLA as measured in skin blister fluid from about 0.2:1 to about 5:1,about 0.5:1 to about 2.5:1 or about 0.6:1 to about 1.5:1.
 16. Thepharmaceutical composition of claim 13, wherein upon oral administrationof multiple daily doses of said composition over a period of about 2 toabout 30 days, each daily dose comprising about 200 mg to about 8000 mgof DGLA, to a fasted subject and to a fed subject, the subjects exhibit:(a) a ratio of free DGLA C. fasted:fed of between 1:1 to 5:1; of about2.5:1, of about 3:1 or of about 3.5:1; (b) a ratio of free DGLA AUC₀₋₂₄fasted:fed of between 1:1 to 5:1; of about 1.5:1,of about 2:1 or ofabout 2.5:1; (c) a ratio of total DGLA C_(max) fasted:fed of between 1:1to 5:1; of about 1:1, of about 1.5:1 or of about 2:1; and/or (d) a ratioof total DGLA AUC₀₋₂₄ fasted:fed of between 1:1 to 5:1; of about 1.5:1,of about 2:1 or of about 2.5:1.
 17. The pharmaceutical composition ofclaim 13 wherein the composition is encapsulated one or more capsuleshells.
 18. The pharmaceutical composition of claim 17, wherein the oneor more capsule shells comprises gelatin and glycerol.
 19. Thepharmaceutical composition of claim 18, wherein up to about 1 g of DGLAor a derivative thereof is encapsulated in each of said one or morecapsule shells.
 20. The pharmaceutical composition of claim 18, whereinabout 200 mg to about 1 g of DGLA or a derivative thereof isencapsulated in each of said one or more capsule shells.
 21. Thepharmaceutical composition of claim 18, wherein the gelatin compriseslime bone gelatin and the one or more capsule shells further comprisesone or more of: glycerol, purified water, titanium dioxide, medium chaintriglycerides and/or lecithin.
 22. The pharmaceutical composition ofclaim 13, wherein the composition comprises not less than 95% by weightDGLA, not more than 5% by weight of any individual fatty acid selectedfrom 20:2ω6, 20:3ω3, 20:4ω6, 20:4ω3 and 20:5ω3, and not more than 2% byweight of unidentified related substances.